ORCID Profile
0000-0002-6415-1639
Current Organisations
University of Sydney
,
University of Melbourne
,
Chris O'Brien Lifehouse
,
University of Wollongong - Innovation Campus
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Research Topics
In Research Link Australia (RLA), "Research Topics" refer to ANZSRC FOR and SEO codes. These topics are either sourced from ANZSRC FOR and SEO codes listed in researchers' related grants or generated by a large language model (LLM) based on their publications.
Top 5 Research Topics
ANZSRC Field of Research (FoR)
Separation Science | Biomaterials | Biomedical Engineering | Analytical Chemistry | Medical Devices | Biochemistry and Cell Biology | Macromolecular and Materials Chemistry | Synthesis of Materials | Manufacturing Engineering not elsewhere classified | Cell Physiology | Receptors and Membrane Biology | Synthetic Biology | Biological Physics | Nanomanufacturing | Functional Materials | Environmental Chemistry (incl. Atmospheric Chemistry) | Analytical Biochemistry
ANZSRC Socio-Economic Objective (SEO)
Expanding Knowledge in the Biological Sciences | Expanding Knowledge in Technology | Expanding Knowledge in the Chemical Sciences | Aquaculture Rock Lobster | Human Diagnostics | Manufacturing not elsewhere classified | Expanding Knowledge in the Medical and Health Sciences | Water Services and Utilities | Horticultural Crops not elsewhere classified | Expanding Knowledge in Engineering |
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Publications
Karyotype of human ES cells during extended culture
Publisher: Springer Science and Business Media LLC
Date: 04-2004
DOI: 10.1038/NBT0404-381
Comparative analysis of group II metabotropic glutamate receptor immunoreactivity in Brodmann's area 46 of the dorsolateral prefrontal cortex from patients with schizophrenia and normal subjects
Publisher: Springer Science and Business Media LLC
Date: 02-2002
Abstract: Glutamate is the primary excitatory neurotransmitter in the mammalian central nervous system, and a key neurotransmitter in prefrontal cortical function. Converging lines of evidence implicate prefrontal cortical dysfunction in the neurobiology of schizophrenia. Thus, aberrant glutamate neurotransmission may underlie schizophrenia and other complex disorders of behavior. Group II metabotropic receptors (mGluRs) are important modulators of glutamatergic and non-glutamatergic neurotransmission. Moreover, in an animal model, an agonist for group II mGluRs has been shown to reverse the behavioral, locomotor, and cognitive effects of the psychotomimetic drug phencyclidine. Accordingly, group II mGluRs constitute attractive targets for the pharmacotherapeutics and study of schizophrenia. Using immunocytochemistry and Western immunoblotting, we compared the localization and levels of group II mGluRs in Brodmann's area 46 of the dorsolateral prefrontal cortex from patients with schizophrenia and normal subjects. Consistent with previous reports, we found that immunolabeling of group II mGluRs is prominent in Brodmann's area 46. The majority of labeling was present on axon terminals distributed in a lamina-specific fashion. No apparent difference in the cellular localization or laminar distribution of immunoreactive group II mGluRs was noted between the two diagnostic groups. Similarly, the levels of receptor immunoreactivity determined by quantitative Western immunoblotting were comparable between schizophrenic patients and normal subjects. We conclude that while the function of group II mGluRs in Brodmann's area 46 of dorsolateral prefrontal cortex may be altered in patients with schizophrenia, this is not evident at the level of protein expression using an antibody against mGluR2 and mGluR3.
Pre-differentiation of human neural stem cells into GABAergic neurons prior to transplant results in greater repopulation of the damaged brain and accelerates functional recovery after transient ischemic stroke
Publisher: Springer Science and Business Media LLC
Date: 29-09-2015
DOI: 10.1186/S13287-015-0175-1
Abstract: Despite attempts to prevent brain injury during the hyperacute phase of stroke, most sufferers end up with significant neuronal loss and functional deficits. The use of cell-based therapies to recover the injured brain offers new hope. In the current study, we employed human neural stem cells (hNSCs) isolated from subventricular zone (SVZ), and directed their differentiation into GABAergic neurons followed by transplantation to ischemic brain. Pre-differentiated GABAergic neurons, undifferentiated SVZ-hNSCs or media alone were stereotaxically transplanted into the rat brain (n=7/group) 7 days after endothelin-1 induced stroke. Neurological outcome was assessed by neurological deficit scores and the cylinder test. Transplanted cell survival, cellular phenotype and maturation were assessed using immunohistochemistry and confocal microscopy. Behavioral assessments revealed accelerated improvements in motor function 7 days post-transplant in rats treated with pre-differentiated GABAergic cells in comparison to media alone and undifferentiated hNSC treated groups. Histopathology 28 days-post transplant indicated that pre-differentiated cells maintained their GABAergic neuronal phenotype, showed evidence of synaptogenesis and up-regulated expression of both GABA and calcium signaling proteins associated with neurotransmission. Rats treated with pre-differentiated cells also showed increased neurogenic activity within the SVZ at 28 days, suggesting an additional trophic role of these GABAergic cells. In contrast, undifferentiated SVZ-hNSCs predominantly differentiated into GFAP-positive astrocytes and appeared to be incorporated into the glial scar. Our study is the first to show enhanced exogenous repopulation of a neuronal phenotype after stroke using techniques aimed at GABAergic cell induction prior to delivery that resulted in accelerated and improved functional recovery.
Schwann cells revert to non-myelinating phenotypes in the deafened rat cochlea
Publisher: Wiley
Date: 25-09-2007
DOI: 10.1111/J.1460-9568.2007.05811.X
Abstract: Loss of sensory hair cells within the cochlea results in a permanent sensorineural hearing loss and initiates the gradual degeneration of spiral ganglion neurons (SGNs) - the primary afferent neurons of the cochlea. While these neurons are normally myelinated via Schwann cells, loss of myelin occurs as a precursor to neural degeneration. However, the relationship between demyelination and the status of Schwann cells in deafness is not well understood. We used a marker of peripheral myelin (myelin protein zero P0) and a marker of Schwann cells (S100) to determine the temporal sequence of myelin and Schwann cell loss as a function of duration of deafness. Rat pups were systemically deafened for periods ranging from 2 weeks to greater than 6 months by co-administration of frusemide and gentamicin. Cochleae were cryosectioned and quantitative immunohistochemistry used to determine the extent of P0 and S100 labelling within the peripheral processes, SGN soma and their central processes within the modiolus. SGN density was also determined for each cochlear turn. P0 labelling decreased throughout the cochlea with increasing duration of deafness. The reduction in P0 labelling occurred at a faster rate than the SGN loss. In contrast, S100 labelling was not significantly reduced compared with age-matched controls in any cochlear region until 6 months post-deafening. These results suggest that Schwann cells may revert to non-myelinating phenotypes in response to deafness and exhibit greater survival traits than SGNs. The potential clinical significance of these findings for cochlear implants is discussed.
G-protein Coupled Receptors in Stem Cell Self-Renewal and Differentiation
Publisher: Springer Science and Business Media LLC
Date: 13-07-2010
DOI: 10.1007/S12015-010-9167-9
Abstract: Stem cells have great potential for understanding early development, treating human disease, tissue trauma and early phase drug discovery. The factors that control the regulation of stem cell survival, proliferation, migration and differentiation are still emerging. Some evidence now exists demonstrating the potent effects of various G-protein coupled receptor (GPCR) ligands on the biology of stem cells. This review aims to give an overview of the current knowledge of the regulation of embryonic and somatic stem cell maintenance and differentiation by GPCR ligands.
Biological Markers and Schizophrenia
Publisher: SAGE Publications
Date: 12-2000
DOI: 10.1080/000486700230
Abstract: The delivery of biological markers for schizophrenia would greatly assist preventative strategies by identifying at-risk in iduals who could then be monitored and treated in a manner with a view to minimising subsequent morbidity. This paper aims to present a selection of biological measures that may indicate risk of schizophrenia. A selective and brief review is provided of intensively studied putative markers, including enlarged cerebral ventricles, dopamine D2 receptor density, hetamine-stimulated central nervous system dopamine release, plasma homovanillic acid and smooth pursuit eye tracking dysfunction. A number of biological measures have been reported to be correlated with schizophrenia. Presently, none of these measures has satisfactory performance characteristics in terms of predictive validity, noninvasiveness, ease of testing and low cost that would enable their widespread use. However, a few have potential for further investigation and development.
A Standard Nomenclature for Referencing and Authentication of Pluripotent Stem Cells
Publisher: Elsevier BV
Date: 2018
Clinically Amendable, Defined, and Rapid Induction of Human Brain Organoids from Induced Pluripotent Stem Cells
Publisher: Springer New York
Date: 2017
DOI: 10.1007/7651_2017_95
Abstract: Human brain organoids provide opportunities to produce three-dimensional (3D) brain-like tissues for biomedical research and translational drug discovery, toxicology, and tissue replacement. Here we describe a protocol for rapid and defined induction of brain organoids from human induced pluripotent stem cells (iPSCs), using commercially available culture and differentiation media and a cheap, easy to handle and clinically approved semisynthetic hydrogel. Importantly, the methodology is uncomplicated, well-defined, and reliable for reproducible and scalable organoid generation, and amendable to principles of current good laboratory practice (cGLP), with the potential for prospective adaptation to current good manufacturing practice (cGMP) toward clinical compliance.
Conducting Polymer Mediated Electrical Stimulation Induces Multilineage Differentiation with Robust Neuronal Fate Determination of Human Induced Pluripotent Stem Cells
Publisher: MDPI AG
Date: 09-03-2020
DOI: 10.3390/CELLS9030658
Abstract: Electrical stimulation is increasingly being used to modulate human cell behaviour for biotechnological research and therapeutics. Electrically conductive polymers (CPs) such as polypyrrole (PPy) are amenable to in vitro and in vivo cell stimulation, being easy to synthesise with different counter ions (dopants) to augment biocompatibility and cell-effects. Extending our earlier work, which showed that CP-mediated electrical stimulation promotes human neural stem cell differentiation, here we report using electroactive PPy containing the anionic dopant dodecylbenzenesulfonate (DBS) to modulate the fate determination of human induced pluripotent stem cells (iPSCs). Remarkably, the stimulation without conventional chemical inducers resulted in the iPSCs differentiating to cells of the three germ lineages—endoderm, ectoderm, and mesoderm. The unstimulated iPSC controls remained undifferentiated. Phenotypic characterisation further showed a robust induction to neuronal fate with electrical stimulation, again without customary chemical inducers. Our findings add to the growing body of evidence supporting the use of electrical stimulation to augment stem cell differentiation, more specifically, pluripotent stem cell differentiation, and especially neuronal induction. Moreover, we have shown the versatility of electroactive PPy as a cell-compatible platform for advanced stem cell research and translation, including identifying novel mechanisms of fate regulation, tissue development, electroceuticals, and regenerative medicine.
Bioengineering Clinically Relevant Cardiomyocytes and Cardiac Tissues from Pluripotent Stem Cells
Publisher: MDPI AG
Date: 16-03-2021
DOI: 10.3390/IJMS22063005
Abstract: The regenerative capacity of cardiomyocytes is insufficient to functionally recover damaged tissue, and as such, ischaemic heart disease forms the largest proportion of cardiovascular associated deaths. Human-induced pluripotent stem cells (hiPSCs) have enormous potential for developing patient specific cardiomyocytes for modelling heart disease, patient-based cardiac toxicity testing and potentially replacement therapy. However, traditional protocols for hiPSC-derived cardiomyocytes yield mixed populations of atrial, ventricular and nodal-like cells with immature cardiac properties. New insights gleaned from embryonic heart development have progressed the precise production of subtype-specific hiPSC-derived cardiomyocytes however, their physiological immaturity severely limits their utility as model systems and their use for drug screening and cell therapy. The long-entrenched challenges in this field are being addressed by innovative bioengingeering technologies that incorporate biophysical, biochemical and more recently biomimetic electrical cues, with the latter having the potential to be used to both direct hiPSC differentiation and augment maturation and the function of derived cardiomyocytes and cardiac tissues by mimicking endogenous electric fields.
The generation of GLP-grade human embryonic stem cell banks from four clinical-grade cell lines for preclinical research
Publisher: Springer Science and Business Media LLC
Date: 23-02-2010
DOI: 10.1007/S11626-010-9287-1
Abstract: The Singapore Stem Cell Bank has generated human embryonic stem cell banks from clinical-grade cell lines ESI-017, ESI-035, ESI-049, and ESI-053. All banks were prepared and characterized according to principles of Good Laboratory Practice for quality assurance. Importantly, each cell line has clearly documented and approved ethical provenance and meets recognized standards for performance and safety. The banks are intended to facilitate the translation of stem cell research to clinical medicine by enabling early phase research and development with high-quality, low-cost cells that are also available as clinical-grade stocks.
Differentiation of stem cells from human infrapatellar fat pad: Characterization of cells undergoing chondrogenesis
Publisher: Mary Ann Liebert Inc
Date: 08-2014
DOI: 10.1089/TEN.TEA.2013.0657
Abstract: Hyaline cartilage repair is a significant challenge in orthopedics and current techniques result in formation of fibrocartilage. Human infrapatellar fat pad (hIPFP)-derived mesenchymal stem cells (MSCs) are capable of differentiation into multiple tissue lineages, including cartilage and bone. Chondrogenesis is a crucial part of normal skeletal development but the molecular mechanisms are yet to be completely defined. In this study we sourced hIPFP-derived MSCs utilizing chondrogenic growth factors, transforming growth factor beta-3, and bone morphogenetic protein-6, to form hyaline-like cartilage in micromass cultures and we studied chondrogenic development of 7, 14, and 28 days. The purpose of this study was (1) to characterize chondrogenesis from MSCs derived from hIPFP tissue by conventional techniques and (2) to characterize temporal changes of key molecular components during chondrogenesis using microarray gene expression. Endpoints included histology, immunohistochemistry (IHC), gene expression profiles using a microarray technique, and changes in expression of specific genes using quantitative real-time polymerase chain reaction. Over 14-28 days, clusters of encapsulated chondrocytes formed surrounded by collagen type II and aggrecan in the extracellular matrix (ECM). Collagen type II and aggrecan production was confirmed using IHC and chondrogenic lineage markers were studied SRY-related transcription factor (SOX9), collagen type II alpha 1 (COL2A1), and aggrecan gene expression increased significantly over the time course. Normalized microarray highlighted 608 differentially expressed genes 10 chondrogenic genes were upregulated (2- to 87-fold), including COL2A1, COL10A1, COL9A1, COL11A1, COL9A2, COL11A2, COL1A1, COMP, SOX9, and COL3A1. We found that the upregulated genes (twofold or greater) represent significant level of expression (enrichment score) for the ECM structural constituent of the molecular functional at days 7, 14, and 28 during chondrogenesis. Therefore, we have successfully demonstrated in vitro production of hyaline-like cartilage from IPFP-derived MSCs in micromass culture. Microarray has provided information concerning genes involved in chondrogenesis of hIPFP-derived MSCs and our approach offers a viable strategy for generating clinically relevant cartilage for therapeutic use.
Muscarinic1 and 2 receptor mRNA in the human caudate-putamen: No change in m1 mRNA in schizophrenia
Publisher: Springer Science and Business Media LLC
Date: 03-2000
Abstract: Studies using tissue obtained at autopsy suggest that changes in cholinergic neurons could be important in the pathology of schizophrenia.1-4 We have previously reported a decrease in [3H]pirenzepine binding5 and [3H]AF-DX 384 binding6 to caudate-putamen (CP) from subjects who had schizophrenia. Under the conditions chosen, [3H]pirenzepine would predominately bind to muscarinic1 (M1) and muscarinic4 (M4) receptors,7whereas [3H]AF-DX 384 would mainly bind to muscarinic2 (M2) and M4 receptors.8 Given the relative concentrations of M1, M2 and M4 receptors in the human CP and the magnitude of the decreases in radioligand binding in schizophrenia, our results most likely reflected a change in the density of M1 and M2 receptors in the CP from the schizophrenic subjects. In situ hybridisation has now been used to determine levels of m1 and m2 mRNA in CP from 14 schizophrenic and 16 control subjects previously used for radioligand binding. m2 mRNA in the CP from the schizophrenic and control subjects was below the sensitivity of in situhybridisation. There was no difference in the levels of m1 mRNA in CP from schizophrenic and control subjects (mean +/- SEM: 103 +/- 16 vs106 +/- 17 fmol [35S]oligonucleotide probe g-1estimated tissue equivalents, P = 0.91). In conclusion, data from our radioligand binding studies show decreases in [3H]pirenzepine binding that are likely to reflect a decrease in the density of M1 receptors in CP from schizophrenic subjects. Our data in this study show the absence of a concomitant change in mRNA coding for that receptor.
Biobanks for Pluripotent Stem Cells
Publisher: Elsevier
Date: 2012
Banking Human Induced Pluripotent Stem Cells: Lessons Learned from Embryonic Stem Cells?
Publisher: Elsevier BV
Date: 10-2013
DOI: 10.1016/J.STEM.2013.09.007
Abstract: The generation of human embryonic stem cell banking networks has ensured that well-characterized and quality controlled stem cell lines are broadly accessible to researchers worldwide. Here, we provide recommendations for engaging these established networks in efforts to build similar resources for the distribution and collection of induced pluripotent stem cells.
The International Stem Cell Banking Initiative (ISCBI): raising standards to bank on
Publisher: Springer Science and Business Media LLC
Date: 03-03-2010
DOI: 10.1007/S11626-010-9301-7
Abstract: The International Stem Cell Banking Initiative (ISCBI) aims to create a global network of stem cell banks to facilitate best practice in stem cell research and clinical cell delivery, primary objectives of national and local governments worldwide and stem cell organizations such the International Stem Cell Forum and the International Society of Stem Cell Research. This paper is a brief overview of ISCBI, its primary activities, potential network participants, and the challenges for harmonizing stem cell banking on a global level.
Microarray analysis of gene expression in the prefrontal cortex in schizophrenia: a preliminary study
Publisher: Elsevier BV
Date: 11-2002
DOI: 10.1016/S0920-9964(01)00377-2
Abstract: Microarray studies can be used to examine expression levels for large numbers of genes simultaneously and may be applied to identify genes involved in schizophrenia. A microarray with 1127 brain-relevant genes was used to screen relative gene expression in the dorsolateral prefrontal cortex (DLPFC) from three pools of patients with schizophrenia (n = 15) and three matched control pools (n = 15). Pooling of tissue s les was employed as a strategy to detect changes in gene expression that are consistently found across in idual cases of schizophrenia. Differences in gene expression were examined by z-ratios in addition to traditional normalized ratios. Three genes that showed consistently decreased expression in schizophrenia by both z-ratio differences and decreased normalized numerical ratios were identified. These were histidine triad nucleotide-binding protein (HINT), ubiquitin conjugating enzyme E2N (UBE2N) and glutamate receptor, ionotropic, AMPA 2 (GRIA2). Moreover, HINT gene expression was decreased to a similar degree in a prior study. In addition, a decrease in AMPA receptor expression is consistent with a decrease in glutamate synaptic function. These results are subject to limitations based on variations inherent to human subjects and tissue s les, possible effects of neuroleptic treatment, and the requirement for verification using independent techniques.
Electrical stimulation using conductive polymer polypyrrole counters reduced neurite outgrowth of primary prefrontal cortical neurons from NRG1-KO and DISC1-LI mice
Publisher: Springer Science and Business Media LLC
Date: 15-02-2017
DOI: 10.1038/SREP42525
Abstract: Deficits in neurite outgrowth, possibly involving dysregulation of risk genes neuregulin-1 (NRG1) and disrupted in schizophrenia 1 (DISC1) have been implicated in psychiatric disorders including schizophrenia. Electrical stimulation using conductive polymers has been shown to stimulate neurite outgrowth of differentiating human neural stem cells. This study investigated the use of the electroactive conductive polymer polypyrrole (Ppy) to counter impaired neurite outgrowth of primary pre-frontal cortical (PFC) neurons from NRG1-knock out (NRG1-KO) and DISC1-locus impairment (DISC1-LI) mice. Whereas NRG1-KO and DISC1-LI exhibited reduced neurite length and number of neurite branches compared to wild-type controls, this was not apparent for cultures on electroactive Ppy. Additionally, the use of the Ppy substrate normalised the synaptophysin and PSD95 protein and mRNA expression whereas both are usually reduced by NRG1-KO or DISC1-LI. Our findings support the utility of Ppy mediated electrical stimulation to prevent the reduction of neurite outgrowth and related synaptic protein expression in the primary PFC neurons from NRG1-KO and DISC1-LI mice, providing proof-of-concept for treating neurodevelopmental diseases including schizophrenia.
A 3D printed graphene electrode device for enhanced and scalable stem cell culture, osteoinduction and tissue building
Publisher: Elsevier BV
Date: 03-2021
From Free Tissue Transfer to Hydrogels: A Brief Review of the Application of the Periosteum in Bone Regeneration
Publisher: MDPI AG
Date: 21-09-2023
DOI: 10.3390/GELS9090768
Molecular interactions and forces of adhesion between single human neural stem cells and gelatin methacrylate hydrogels of varying stiffness
Publisher: Elsevier BV
Date: 04-2020
Electrical Stimulation Using Conductive Polymer Polypyrrole Promotes Differentiation of Human Neural Stem Cells: A Biocompatible Platform for Translational Neural Tissue Engineering
Publisher: Mary Ann Liebert Inc
Date: 04-2015
Wet-Spun Trojan Horse Cell Constructs for Engineering Muscle.
Publisher: Frontiers Media SA
Date: 20-02-2020
Cochlear immunochemistry - a new technique based on gelatin embedding
Publisher: Elsevier BV
Date: 10-2003
DOI: 10.1016/S0165-0270(03)00211-5
Abstract: Histological processing of the cochlea for immunochemistry is often a compromise between good anatomical resolution and preservation of antigenicity. Techniques able to preserve tissue architecture invariably demand elevated temperatures and harsh chemicals or a combination of both. The likely result is reduced antigenicity, enzyme activity and nucleic acid integrity. We have modified an existing embedding medium for use in the cochlea that operates at physiological temperature and avoids denaturing agents and organic solvents. Tissue antigenicity is maximised and anatomical detail preserved, normally two mutually exclusive goals. The method is attractive because of its simplicity, speed and transparency for easy cochlear orientation. It is also likely to be adaptable for the infiltration of other heterogeneous structures prone to distortion during frozen sectioning.
Cell Processing for 3D Bioprinting: Quality Requirements for Quality Assurance in Fundamental Research and Translation
Publisher: Springer US
Date: 2020
DOI: 10.1007/978-1-0716-0520-2_2
Abstract: Bioprinting is an additive manufacturing process where biomaterials-based inks are printed layer-by-layer to create three-dimensional (3D) structures that mimic natural tissues. Quality assurance for 3D bioprinting is paramount to undertaking fundamental research and preclinical and clinical product development. It forms part of quality management and is vital to reproducible and safe tissue fabrication, function, and regulatory approval for translational application. This chapter seeks to place the implementation of quality practices in 3D bioprinting front-of-mind, with emphasis on cell processing, although important to all components and procedures of the printing pipeline.
Biomaterials for corneal bioengineering
Publisher: IOP Publishing
Date: 06-03-2018
Associations between socioeconomic status and environmental toxicant concentrations in adults in the USA: NHANES 2001-2010.
Publisher: Elsevier BV
Date: 09-2013
DOI: 10.1016/J.ENVINT.2013.06.017
Abstract: Low level chronic exposure to toxicants is associated with a range of adverse health effects. Understanding the various factors that influence the chemical burden of an in idual is of critical importance to public health strategies. We investigated the relationships between socioeconomic status (SES) and bio-monitored chemical concentration in five cross-sectional waves of the U.S. National Health and Nutrition Examination Survey (NHANES). We utilised adjusted linear regression models to investigate the association between 179 toxicants and the poverty income ratio (PIR) for five NHANES waves. We then selected a subset of chemicals associated with PIR in 3 or more NHANES waves and investigated potential mediating factors using structural equation modelling. PIR was associated with 18 chemicals in 3 or more NHANES waves. Higher SES in iduals had higher burdens of serum and urinary mercury, arsenic, caesium, thallium, perfluorooctanoic acid, perfluorononanoic acid, mono(carboxyoctyl) phthalate and benzophenone-3. Inverse associations were noted between PIR and serum and urinary lead and cadmium, antimony, bisphenol A and three phthalates (mono-benzyl, mono-isobutyl, mono-n-butyl). Key mediators included fish and shellfish consumption for the PIR, mercury, arsenic, thallium and perfluorononanoic acid associations. Sunscreen use was an important mediator in the benzophenone-3/PIR relationship. The association between PIR and cadmium or lead was partially mediated by smoking, occupation and diet. These results provide a comprehensive analysis of exposure patterns as a function of socioeconomic status in US adults, providing important information to guide future public health remediation measures to decrease toxicant and disease burdens within society.
Development of a porous 3D graphene-PDMS scaffold for improved osseointegration
Publisher: Elsevier BV
Date: 11-2017
DOI: 10.1016/J.COLSURFB.2017.07.087
Abstract: Osseointegration in orthopedic surgery plays an important role for bone implantation success. Traditional treatment of implant surface aimed at improved osseointegration has limited capability for its poor performance in supporting cell growth and proliferation. Polydimethylsiloxane (PDMS) is a widely used silicon-based organic polymer material with properties that are useful in cosmetics, domestic applications and mechanical engineering. In addition, the biocompatibility of PDMS, in part due to the high solubility of oxygen, makes it an ideal material for cell-based implants. Notwithstanding its potential, a property that can inhibit PDMS bioactivity is the high hydrophobicity, limiting its use to date in tissue engineering. Here, we describe an efficient approach to produce porous, durable and cytocompatible PDMS-based 3D structures, coated with reduced graphene oxide (RGO). The RGO/PDMS scaffold has good mechanical strength and with pore sizes ranging from 10 to 600μm. Importantly, the scaffold is able to support growth and differentiation of human adipose stem cells (ADSCs) to an osteogenic cell lineage, indicative of its potential as a transition structure of an osseointegrated implant.
Conductive Tough Hydrogel for Bioapplications
Publisher: Wiley
Date: 13-12-2018
Abstract: Biocompatible conductive tough hydrogels represent a new class of advanced materials combining the properties of tough hydrogels and biocompatible conductors. Here, a simple method, to achieve a self-assembled tough elastomeric composite structure that is biocompatible, conductive, and with high flexibility, is reported. The hydrogel comprises polyether-based liner polyurethane (PU), poly(3,4-ethylenedioxythiophene) (PEDOT) doped with poly(4-styrenesulfonate) (PSS), and liquid crystal graphene oxide (LCGO). The polyurethane hybrid composite (PUHC) containing the PEDOT:PSS, LCGO, and PU has a higher electrical conductivity (10×), tensile modulus (>1.6×), and yield strength (>1.56×) compared to respective control s les. Furthermore, the PUHC is biocompatible and can support human neural stem cell (NSC) growth and differentiation to neurons and supporting neuroglia. Moreover, the stimulation of PUHC enhances NSC differentiation with enhanced neuritogenesis compared to unstimulated cultures. A model describing the synergistic effects of the PUHC components and their influence on the uniformity, biocompatibility, and electromechanical properties of the hydrogel is presented.
Ex Vivo Preservation of Ovine Periosteum Using a Perfusion Bioreactor System
Publisher: MDPI AG
Date: 27-06-2023
Abstract: Periosteum is a highly vascularized membrane lining the surface of bones. It plays essential roles in bone repair following injury and reconstruction following invasive surgeries. To broaden the use of periosteum, including for augmenting in vitro bone engineering and/or in vivo bone repair, we have developed an ex vivo perfusion bioreactor system to maintain the cellular viability and metabolism of surgically resected periosteal flaps. Each specimen was placed in a 3D printed bioreactor connected to a peristaltic pump designed for the optimal flow rates of tissue perfusate. Nutrients and oxygen were perfused via the periosteal arteries to mimic physiological conditions. Biochemical assays and histological staining indicate component cell viability after perfusion for almost 4 weeks. Our work provides the proof-of-concept of ex vivo periosteum perfusion for long-term tissue preservation, paving the way for innovative bone engineering approaches that use autotransplanted periosteum to enhance in vivo bone repair.
Chronic depolarization enhances the trophic effects of brain-derived neurotrophic factor in rescuing auditory neurons following a sensorineural hearing loss
Publisher: Wiley
Date: 2005
DOI: 10.1002/CNE.20564
Expansion of Undifferentiated Human Embryonic Stem Cells
Publisher: Springer Netherlands
Date: 2005
3D graphene-containing structures for tissue engineering
Publisher: Elsevier BV
Date: 12-2019
Human Embryonic Stem Cell Therapies for Neurodegenerative Diseases
Publisher: Bentham Science Publishers Ltd.
Date: 06-2011
DOI: 10.2174/187152711795564001
Abstract: There is a renewed enthusiasm for the clinical translation of human embryonic stem (hES) cells. This is abetted by putative clinically-compliant strategies for hES cell maintenance and directed differentiation, greater understanding of and accessibility to cells through formal cell registries and centralized cell banking for distribution, the revised US government policy on funding hES cell research, and paradoxically the discovery of induced pluripotent stem (iPS) cells. Additionally, as we consider the constraints (practical and fiscal) of delivering cell therapies for global healthcare, the more efficient and economical application of allogeneic vs autologous treatments will bolster the clinical entry of hES cell derivatives. Neurodegenerative disorders such as Parkinson's disease are primary candidates for hES cell therapy, although there are significant hurdles to be overcome. The present review considers key advances and challenges to translating hES cells into novel therapies for neurodegenerative diseases, with special consideration given to Parkinson's disease and Alzheimer's disease. Importantly, despite the focus on degenerative brain disorders and hES cells, many of the issues canvassed by this review are relevant to systemic application of hES cells and other pluripotent stem cells such as iPS cells.
Human stem cells for modeling neurological disorders: Accelerating the drug discovery pipeline
Publisher: Wiley
Date: 03-11-2008
DOI: 10.1002/JCB.21967
Abstract: The availability of human stem cells heralds a new era for modeling normal and pathologic tissues and developing therapeutics. For ex le, the in vitro recapitulation of normal and aberrant neurogenesis holds significant promise as a tool for de novo modeling of neurodevelopmental and neurodegenerative diseases. Translational applications include deciphering brain development, function, pathologies, traditional medications, and drug discovery for novel pharmacotherapeutics. For the latter, human stem cell-based assays represent a physiologically relevant and high-throughput means to assess toxicity and other undesirable effects early in the drug development pipeline, avoiding late-stage attrition whilst expediting proof-of-concept of genuine drug candidates. Here we consider the potential of human embryonic, adult, and induced pluripotent stem cells for studying neurological disorders and preclinical drug development.
Repressed by a NuRD
Publisher: Springer Science and Business Media LLC
Date: 03-2006
DOI: 10.1038/NCB0306-212
Cryobanking Pluripotent Stem Cells
Publisher: Springer New York
Date: 2017
DOI: 10.1007/978-1-4939-6921-0_11
Abstract: Cryobanking human pluripotent stem cells (hPSCs), be they human embryonic (hESCs) or induced pluripotent stem cells (iPSCs), is essential for their use in research and cell-based therapeutics. Working and master cell banks can be generated with a desired level of quality assurance applied during cell freezing and storage. Conventional vitrification has evolved to more advanced control rate freezing, culminating in a myriad of published protocols with variable proficiencies and clinical efficacies. Notwithstanding, standardized and reliable protocols are necessary for basic science through to applied research and clinical product development. This chapter details several methods for hPSC cryopreservation, suitable for routine application, high-quality research, and adaptable for clinical compliance.
3D bioprinting human induced pluripotent stem cell constructs for in situ cell proliferation and successive multilineage differentiation
Publisher: Wiley
Date: 24-05-2017
Abstract: The ability to create 3D tissues from induced pluripotent stem cells (iPSCs) is poised to revolutionize stem cell research and regenerative medicine, including in idualized, patient-specific stem cell-based treatments. There are, however, few ex les of tissue engineering using iPSCs. Their culture and differentiation is predominantly planar for monolayer cell support or induction of self-organizing embryoids (EBs) and organoids. Bioprinting iPSCs with advanced biomaterials promises to augment efforts to develop 3D tissues, ideally comprising direct-write printing of cells for encapsulation, proliferation, and differentiation. Here, such a method, employing a clinically amenable polysaccharide-based bioink, is described as the first ex le of bioprinting human iPSCs for in situ expansion and sequential differentiation. Specifically, we have extrusion printed the bioink including iPSCs, alginate (Al 5% weight/volume [w/v]), carboxymethyl-chitosan (5% w/v), and agarose (Ag 1.5% w/v), crosslinked the bioink in calcium chloride for a stable and porous construct, proliferated the iPSCs within the construct and differentiated the same iPSCs into either EBs comprising cells of three germ lineages-endoderm, ectoderm, and mesoderm, or more homogeneous neural tissues containing functional migrating neurons and neuroglia. This defined, scalable, and versatile platform is envisaged being useful in iPSC research and translation for pharmaceuticals development and regenerative medicine.
Low muscarinic receptor binding in prefrontal cortex from subjects with schizophrenia: A study of brodmann's areas 8, 9, 10, and 46 and the effects of neuroleptic drug treatment
Publisher: American Psychiatric Association Publishing
Date: 06-2001
DOI: 10.1176/APPI.AJP.158.6.918
Abstract: Aberrant cholinergic inputs and synaptic neurotransmission in the prefrontal cortex induce cognitive impairment, which is a central feature of schizophrenia. Postsynaptic excitatory muscarinic cholinergic M(1) and M(4) receptors are the major cholinoceptive targets in the prefrontal cortex and hence may be involved in the pathology and/or pharmacotherapeutics of schizophrenia. Using quantitative autoradiography, the authors analyzed the binding of the M(1)/M(4) receptor selective antagonist [(3)H]pirenzepine in prefrontal cortex (Brodmann's areas 8, 9, 10, and 46) from schizophrenia patients who had (N=6) or had not (N=11) been treated with the anticholinergic agent benztropine mesylate and from normal comparison subjects (N=20). Moreover, preliminary studies of [(3)H]pirenzepine binding in rat frontal cortex following administration of antipsychotic drugs or benztropine mesylate were performed. Relative to those of comparison subjects, the mean levels of [(3)H]pirenzepine binding were significantly lower in Brodmann's areas 9 and 46 of the schizophrenia patients not treated with benztropine mesylate (18% lower in Brodmann's area 9 and 21% lower in Brodmann's area 46) and in all four examined regions of the patients who had received benztropine (51%-64% lower). Antipsychotic or anticholinergic drugs tended to increase or have no effect on the density of [(3)H]pirenzepine-labeled receptors in rat frontal cortex. Because M(1) and M(4) receptors are critical to the functions of prefrontal cortical acetylcholine, the present findings suggest a functional impairment in cholinergic neurotransmission in schizophrenia and the possibility that muscarinic receptors are involved in the pharmacotherapeutics of the disorder.
Differentiation of Pluripotent Stem Cells: An Overview
Publisher: Wiley
Date: 29-11-2010
Setting Quality Standards for Stem Cell Banking, Research and Translation: The International Stem Cell Banking Initiative
Publisher: Springer New York
Date: 2014
Culturing and Cryobanking Human Neural Stem Cells
Publisher: Springer New York
Date: 2017
DOI: 10.1007/978-1-4939-6921-0_15
Abstract: The discovery and study of human neural stem cells has advanced our understanding of human neurogenesis, and the development of novel therapeutics based on neural cell replacement. Here, we describe methods to culture and cryopreserve human neural stem cells (hNSCs) for expansion and banking. Importantly, the protocols ensure that the multipotency of hNSCs is preserved to enable differentiation to neurons and supporting neuroglia.
Fate of embryonic stem cells transplanted into the deafened mammalian cochlea
Publisher: SAGE Publications
Date: 05-2006
DOI: 10.3727/000000006783981819
Abstract: Spiral ganglion neurons (SGNs), the primary afferent neurons of the cochlea, degenerate following a sensorineural hearing loss (SNHL) due to lack of trophic support normally received from hair cells. Cell transplantation is emerging as a potential strategy for inner ear rehabilitation, as injected cells may be able to replace damaged SGNs in the deafened cochlea. An increase in the number of surviving SGNs may result in improved efficacy of cochlear implants (CIs). We examined the survival of partially differentiated mouse embryonic stem cells (MESCs), following xenograft transplantation into the deafened guinea pig cochlea (n = 15). Cells were delivered directly into the left scala tympani via microinjection through the round window. Small numbers of MESCs were detected in the scala tympani for up to 4 weeks following transplantation and a proportion of these cells retained expression of neurofilament protein 68 kDa in vivo. While this delivery method requires refinement for effective long-term replacement of damaged SGNs, small numbers of MESCs were capable of survival in the deafened mammalian cochlea for up to 4 weeks, without causing an inflammatory tissue response.
Human Neural Tissues from Neural Stem Cells Using Conductive Biogel and Printed Polymer Microelectrode Arrays for 3D Electrical Stimulation
Publisher: Wiley
Date: 06-06-2019
Abstract: Electricity is important in the physiology and development of human tissues such as embryonic and fetal development, and tissue regeneration for wound healing. Accordingly, electrical stimulation (ES) is increasingly being applied to influence cell behavior and function for a biomimetic approach to in vitro cell culture and tissue engineering. Here, the application of conductive polymer (CP) poly(3,4-ethylenedioxythiophene)-polystyrenesulfonate (PEDOT:PSS) pillars is described, direct-write printed in an array format, for 3D ES of maturing neural tissues that are derived from human neural stem cells (NSCs). NSCs are initially encapsulated within a conductive polysaccharide-based biogel interfaced with the CP pillar microelectrode arrays (MEAs), followed by differentiation in situ to neurons and supporting neuroglia during stimulation. Electrochemical properties of the pillar electrodes and the biogel support their electrical performance. Remarkably, stimulated constructs are characterized by widespread tracts of high-density mature neurons and enhanced maturation of functional neural networks. Formation of tissues using the 3D MEAs substantiates the platform for advanced clinically relevant neural tissue induction, with the system likely amendable to erse cell types to create other neural and non-neural tissues. The platform may be useful for both research and translation, including modeling tissue development, function and dysfunction, electroceuticals, drug screening, and regenerative medicine.
Defined, Simplified, Scalable, and Clinically Compatible Hydrogel-Based Production of Human Brain Organoids
Publisher: MDPI AG
Date: 11-01-2023
Abstract: Human brain organoids present a new paradigm for modeling human brain organogenesis, providing unprecedented insight to the molecular and cellular processes of brain development and maturation. Other potential applications include in vitro models of disease and tissue trauma, as well as three-dimensional (3D) clinically relevant tissues for pharmaceuticals development and cell or tissue replacement. A key requirement for this emerging technology in both research and medicine is the simple, scalable, and reproducible generation of organoids using reliable, economical, and high-throughput culture platforms. Here we describe such a platform using a defined, clinically compliant, and readily available hydrogel generated from gelatin methacrylate (GelMA). We demonstrate the efficient production of organoids on GelMA from human induced pluripotent stem cells (iPSCs), with scalable production attained using 3D printed GelMA-based multiwell arrays. The differentiation of iPSCs was systematic, rapid, and direct to enable iPSCs to form organoids in their original position following seeding on GelMA, thereby avoiding further cell and organoid disruption. Early neural precursors formed by day 5, neural rosettes and early-stage neurons by day 14, and organoids with cellular and regional heterogeneity, including mature and electrophysiologically active neurons, by day 28. The optimised method provides a simplified and well-defined platform for both research and translation of iPSCs and derivative brain organoids, enabling reliable 3D in vitro modelling and experimentation, as well as the provision of clinically relevant cells and tissues for future therapeutics.
A Comparison of In Vivo Bone Tissue Generation Using Calcium Phosphate Bone Substitutes in a Novel 3D Printed Four-Chamber Periosteal Bioreactor
Publisher: MDPI AG
Date: 21-10-2023
3D Bioprinting Electrically Conductive Bioink with Human Neural Stem Cells for Human Neural Tissues
Publisher: Springer US
Date: 2020
DOI: 10.1007/978-1-0716-0520-2_10
Abstract: Bioprinting cells with an electrically conductive bioink provides an opportunity to produce three-dimensional (3D) cell-laden constructs with the option of electrically stimulating cells in situ during and after tissue development. We and others have demonstrated the use of electrical stimulation (ES) to influence cell behavior and function for a more biomimetic approach to tissue engineering. Here, we detail a previously published method for 3D printing an electrically conductive bioink with human neural stem cells (hNSCs) that are subsequently differentiated. The differentiated tissue constructs comprise functional neurons and supporting neuroglia and are amenable to ES for the purposeful modulation of neural activity. Importantly, the method could be adapted to fabricate and stimulate neural and nonneural tissues from other cell types, with the potential to be applied for both research- and clinical-product development.
Human Brain Organoids and Consciousness: Moral Claims and Epistemic Uncertainty
Publisher: MDPI AG
Date: 07-02-2023
Abstract: Human brain organoids provide a remarkable opportunity to model prenatal human brain biology in vitro by recapitulating features of in utero molecular, cellular and systems biology. An ethical concern peculiar to human brain organoids is whether they are or could become capable of supporting sentience through the experience of pain or pleasure and/or consciousness, including higher cognitive abilities such as self-awareness. Identifying the presence of these traits is complicated by several factors, beginning with consciousness—which is a highly contested concept among neuroscientists, cognitive scientists, and philosophers and so there is no agreed definition. Secondly, given human brain organoids are disembodied, there is no practical way to identify evidence of consciousness as we might in humans or animals. What would count as evidence of organoid consciousness is an emerging area of research. To address concerns about consciousness and human brain organoids, in this paper we clarify the morally relevant aspects of human consciousness, phenomenal experience and embodied development and explore the empirical basis of consciousness to develop a defensible framework for informed decision-making on the moral significance and utility of brain organoids, which can also guide regulation and future research of these novel biological systems.
Decreased muscarinic receptor binding in subjects with schizophrenia: A study of the human hippocampal formation
Publisher: Elsevier BV
Date: 09-2000
DOI: 10.1016/S0006-3223(00)00918-5
Abstract: Acetylcholine is important to hippoc al function, including the processes of learning and memory. Patients with schizophrenia show impaired learning and memory and hippoc al dysfunction. Thus, acetylcholinergic systems may be primarily or secondarily disrupted in the hippoc al formation of schizophrenic patients. The present study tested the hypothesis that [(3)H]pirenzepine-labeled muscarinic cholinergic receptor levels are altered in the hippoc al formation of patients with schizophrenia. We have used quantitative autoradiography to measure [(3)H]pirenzepine binding to M(1) and M(4) receptors in the hippoc al formation from 15 schizophrenic and 18 nonschizophrenic subjects. The mean density of [(3)H]pirenzepine binding was reduced in all regions studied, including the dentate gyrus, sub isions of Ammon's Horn (CA1-CA4), subiculum, and the parahippoc al gyrus, of the schizophrenic cohort. Moreover, unlike controls, there was no significant variation between the mean levels of [(3)H]pirenzepine binding across the subregions of the hippoc al formation from schizophrenic subjects. These findings provide support for a possible involvement of the muscarinic cholinergic system in the pathology and/or treatment of schizophrenia.
Points to consider in the development of seed stocks of pluripotent stem cells for clinical applications: International Stem Cell Banking Initiative (ISCBI)
Publisher: Future Medicine Ltd
Date: 03-2015
DOI: 10.2217/RME.14.93
Metabotropic Glutamate Receptor 2 and 3 Gene Expression in The Human Prefrontal Cortex and Mesencephalon in Schizophrenia
Publisher: Informa UK Limited
Date: 2008
A Report from a Workshop of the International Stem Cell Banking Initiative, Held in Collaboration of Global Alliance for iPSC Therapies and the Harvard Stem Cell Institute, Boston, 2017
Publisher: Oxford University Press (OUP)
Date: 06-08-2019
DOI: 10.1002/STEM.3003
Abstract: This report summarizes the recent activity of the International Stem Cell Banking Initiative held at Harvard Stem Cell Institute, Boston, MA, USA, on June 18, 2017. In this meeting, we aimed to find consensus on ongoing issues of quality control (QC), safety, and efficacy of human pluripotent stem cell banks and their derivative cell therapy products for the global harmonization. In particular, assays for the QC testing such as pluripotency assays test and general QC testing criteria were intensively discussed. Moreover, the recent activities of global stem cell banking centers and the regulatory bodies were briefly summarized to provide an overview on global developments and issues. Stem Cells 2019 :1130–1135
Efficient expansion of clinical-grade human fibroblasts on microcarriers: Cells suitable for ex vivo expansion of clinical-grade hESCs
Publisher: Elsevier BV
Date: 20-03-2008
DOI: 10.1016/J.JBIOTEC.2007.12.007
Abstract: Human embryonic stem cells hold considerable potential for cell-based treatments of a variety of degenerative diseases, including diabetes, ischemic heart failure, and Parkinson's disease. However, advancing research to provide clinical-grade product requires scale-up to therapeutic quantities of stem cells and their differentiated progeny. Most human embryonic stem cell culture platforms require direct support by a fibroblast feeder layer or indirect support using fibroblast conditioned medium. Accordingly, large numbers of clinically compliant fibroblasts will be requisite for stem cell production. Published platforms for feeder production are insufficient for stem cell scale-up, being costly to operate and requiring considerable effort to prepare, maintain and harvest. Here we describe the expansion of cGMP-grade, FDA-approved human foreskin fibroblasts using cGMP-grade reagents and polystyrene-based cationic trimethyl ammonium-coated microcarriers in spinner flasks. Fibroblasts attach rapidly to the microcarriers (T(1/2)=75 min), and expand with a maximum doubling time of 22.5h. Importantly, microcarrier-expanded fibroblasts and their conditioned medium support pluripotent stem cell growth through >5 passages, enabling extended self-renewal and expansion while retaining full differentiation potential. In summary, the method described is an economical and cGMP-compliant means of producing human fibroblast cells in support of cGMP human embryonic stem cell culture.
Smart graphene-cellulose paper for 2D or 3D “origami-inspired” human stem cell support and differentiation
Publisher: Elsevier BV
Date: 04-2019
DOI: 10.1016/J.COLSURFB.2018.12.040
Abstract: Graphene-based materials represent advanced platforms for tissue engineering and implantable medical devices. From a clinical standpoint, it is essential that these materials are produced using non-toxic and non-hazardous methods, and have predictable properties and reliable performance under variable physiological conditions especially when used with a cellular component. Here we describe such a biomaterial, namely smart graphene-cellulose (G-C) paper, and its suitability for traditional planar two-dimensional (2D) or three-dimensional (3D) human cell support, verified by adipose-derived stem cell (ADSC) culture and osteogenic differentiation. G-C paper is prepared using commercially available cellulose tissue paper as a substrate that is coated by immersion-deposition with graphene oxide (GO) followed by reduction to reduced graphene oxide (RGO) without the use of toxic organic solvents. The fabrication process is amenable to large scale production and the resultant papers have low electrical resistivity (up to ∼300 Ω/sq). Importantly, G-C papers can be configured to 3D constructs by lamination with alginate and further modified by folding and rolling for 3D "origami-inspired" cell-laden structures.
Derivation of Human Embryonic Stem Cells
Publisher: Springer New York
Date: 2017
DOI: 10.1007/978-1-4939-6921-0_8
Abstract: Embryonic stem cells (ESCs) represent a mainstay for pluripotent stem cell research and development (R&D) and provide tangible opportunities for clinical translation including cell therapies and drug discovery. Moreover, in spite of the discovery of induced pluripotent stem cells (iPSCs), ESCs are an essential reference point, against which other pluripotent cells are compared. Hence, there is an ongoing need to derive and bank quality-controlled research-grade and clinical-grade ESC lines using established and standardized methods. Here, we provide a concise, step-by-step protocol for the derivation of ESCs from human embryos. While largely based on previously reported method for clinical-grade human ESC (hESC) line derivation, the protocol is suitable for routine application, although adaptable for clinical-compliance.
Development of drug-loaded polymer microcapsules for treatment of epilepsy
Publisher: Royal Society of Chemistry (RSC)
Date: 2017
DOI: 10.1039/C7BM00623C
Abstract: Fibre- and sphere-based microcapsules have been developed, exhibiting controllable uniform morphologies, predictable drug release profiles, and neuro-cytocompatibility.
Toward Clinical Application of Stem Cells for Cardiac Regeneration
Publisher: Elsevier BV
Date: 03-2011
DOI: 10.1016/J.HLC.2010.06.661
Abstract: Heart failure affects more than 10% of the Australian population over age 65, and the ageing population will ensure continued growth of this significant problem. There are various treatment options available, but the growing field of regenerative therapy offers promise to restore or replace tissue lost in those with either congenital or acquired cardiac defects. Stem cells have many potential properties, but they need multiple discussed qualities to succeed in this field such as ease of harvest and multiplication, and most importantly minimal ethical concerns. There are multiple cell types available and one of the challenges will be to find the most appropriate cell type for cardiac regeneration. Cardiac tissue engineering is being explored using both in vitro and in vivo techniques. In vitro methods are primarily limited in terms of the vascularisation and size of the construct. In vivo engineered constructs overcome these limitations in early models, but they are still not ready for human trials. This review aims to provide the reader with an outline of the cell-based and tissue engineering therapies currently being used and developed for cardiac regeneration, as well as some insight into the potential problems that may h er its progress in the future.
Attachment and growth of human embryonic stem cells on microcarriers
Publisher: Elsevier BV
Date: 11-2008
DOI: 10.1016/J.JBIOTEC.2008.07.1997
Abstract: The use of human embryonic stem cells (hESCs) for cell-based therapies will require large quantities of genetically stable pluripotent cells and their differentiated progeny. Traditional hESC propagation entails adherent culture and is sensitive to enzymatic dissociation. These constraints h er modifying method from 2-dimensional flat-bed culture, which is expensive and impractical for bulk cell production. Large-scale culture for clinical use will require innovations such as suspension culture for bioprocessing. Here we describe the attachment and growth kinetics of both murine embryonic stem cells (mESCs) and hESCs on trimethyl ammonium-coated polystyrene microcarriers for feeder-free, 3-dimensional suspension culture. mESCs adhered and expanded according to standard growth kinetics. For hESC studies, we tested aggregate (collagenase-dissociated) and single-cell (TrypLE-dissociated) culture. Cells attached rapidly to beads followed by proliferation. Single-cell cultures expanded 3-fold over approximately 5 days, slightly exceeding that of hESC aggregates. Importantly, single-cell cultures were maintained through 6 passages with a 14-fold increase in cell number while still expressing the undifferentiated markers Oct-4 and Tra 1-81. Finally, hESCs retained their capacity to differentiate towards pancreatic, neuronal, and cardiomyocyte lineages. Our studies provide proof-of-principle of suspension-based expansion of hESCs on microcarriers, as a novel, economical and practical feeder-free means of bulk hESC production.
Corrigendum to “Attachment and growth of human embryonic stem cells on microcarriers” [J. Biotechnol. 138 (2008) 24–32]
Publisher: Elsevier BV
Date: 2009
Netrin-1 as a guidance molecule in the postnatal rat cochlea
Publisher: Elsevier BV
Date: 2005
DOI: 10.1016/J.HEARES.2004.07.004
Abstract: During synaptogenesis a number of growth factors and peptides control the guidance of auditory neuron (spiral ganglion neuron, SGN) axons to their target cells. Furthermore, evidence suggests that these factors exert their actions at discrete times and sites during development. This study demonstrates that the guidance molecule netrin-1 is expressed in the early postnatal rat cochlea, but shows decreasing expression with increasing age. These results suggest that netrin-1 may be involved in guiding axonal growth from SGNs for the onset of innervation, but is not required for maintenance of synaptic connections.
Induced pluripotent stem cells for 2D and 3D modelling the biological basis of schizophrenia and screening possible therapeutics
Publisher: Elsevier BV
Date: 10-2021
DOI: 10.1016/J.BRAINRESBULL.2021.07.004
Abstract: Induced pluripotent stem cells (iPSCs) are providing unprecedented insight into complex neuropsychiatric disorders such as schizophrenia (SZ). Here we review the use of iPSCs for investigating the etiopathology and treatment of SZ, beginning with conventional in vitro two-dimensional (2D monolayer) cell modelling, through to more advanced 3D tissue studies. With the advent of 3D modelling, utilising advanced differentiation paradigms and additive manufacturing technologies, inclusive of patient-specific cerebral/neural organoids and bioprinted neural tissues, such live disease-relevant tissue systems better recapitulate "within-body" tissue function and pathobiology. We posit that by enabling better understanding of biological causality, these evolving strategies will yield novel therapeutic targets and accordingly, drug candidates.
Report of the International Stem Cell Banking Initiative Workshop Activity: Current Hurdles and Progress in Seed-Stock Banking of Human Pluripotent Stem Cells
Publisher: Oxford University Press (OUP)
Date: 24-10-2017
DOI: 10.1002/SCTM.17-0144
Abstract: This article summarizes the recent activity of the International Stem Cell Banking Initiative (ISCBI) held at the California Institute for Regenerative Medicine (CIRM) in California (June 26, 2016) and the Korean National Institutes for Health in Korea (October 19–20, 2016). Through the workshops, ISCBI is endeavoring to support a new paradigm for human medicine using pluripotent stem cells (hPSC) for cell therapies. Priority considerations for ISCBI include ensuring the safety and efficacy of a final cell therapy product and quality assured source materials, such as stem cells and primary donor cells. To these ends, ISCBI aims to promote global harmonization on quality and safety control of stem cells for research and the development of starting materials for cell therapies, with regular workshops involving hPSC banking centers, biologists, and regulatory bodies. Here, we provide a brief overview of two such recent activities, with summaries of key issues raised.
Considerations for modelling diffuse high-grade gliomas and developing clinically relevant therapies
Publisher: Springer Science and Business Media LLC
Date: 04-2023
DOI: 10.1007/S10555-023-10100-7
Abstract: Diffuse high-grade gliomas contain some of the most dangerous human cancers that lack curative treatment options. The recent molecular stratification of gliomas by the World Health Organisation in 2021 is expected to improve outcomes for patients in neuro-oncology through the development of treatments targeted to specific tumour types. Despite this promise, research is hindered by the lack of preclinical modelling platforms capable of recapitulating the heterogeneity and cellular phenotypes of tumours residing in their native human brain microenvironment. The microenvironment provides cues to subsets of glioma cells that influence proliferation, survival, and gene expression, thus altering susceptibility to therapeutic intervention. As such, conventional in vitro cellular models poorly reflect the varied responses to chemotherapy and radiotherapy seen in these erse cellular states that differ in transcriptional profile and differentiation status. In an effort to improve the relevance of traditional modelling platforms, recent attention has focused on human pluripotent stem cell-based and tissue engineering techniques, such as three-dimensional (3D) bioprinting and microfluidic devices. The proper application of these exciting new technologies with consideration of tumour heterogeneity and microenvironmental interactions holds potential to develop more applicable models and clinically relevant therapies. In doing so, we will have a better chance of translating preclinical research findings to patient populations, thereby addressing the current derisory oncology clinical trial success rate.
Pluripotent Human Stem Cells
Publisher: Springer Science and Business Media LLC
Date: 04-2010
DOI: 10.2165/11532270-000000000-00000
Abstract: The need for new and improved pharmacotherapies in medicine, high late-stage compound attrition in drug discovery, and upcoming patent expirations is driving interest by the pharmaceutical industry in pluripotent stem cells for in vitro modeling and early-stage testing of toxicity and target engagement. In particular, human embryonic and induced pluripotent stem cells represent potentially cost-effective and accessible sources of organ-specific cells that foretell in vivo human tissue response to new chemical entities. Here we consider the potential of these cells as novel tools for drug development, including toxicity screening and metabolic profiling. We hold that despite various challenges to translating proof-of-concept screening platforms to industrial use, the promise of research is considerable, and close to being realized.
Three-dimensional neural cultures produce networks that mimic native brain activity
Publisher: Hindawi Limited
Date: 12-12-2018
DOI: 10.1002/TERM.2508
Abstract: Development of brain function is critically dependent on neuronal networks organized through three dimensions. Culture of central nervous system neurons has traditionally been limited to two dimensions, restricting growth patterns and network formation to a single plane. Here, with the use of multichannel extracellular microelectrode arrays, we demonstrate that neurons cultured in a true three-dimensional environment recapitulate native neuronal network formation and produce functional outcomes more akin to in vivo neuronal network activity.
Modelling disrupted-in-schizophrenia 1 loss of function in human neural progenitor cells: tools for molecular studies of human neurodevelopment and neuropsychiatric disorders
Publisher: Springer Science and Business Media LLC
Date: 15-12-2009
DOI: 10.1038/MP.2009.131
Variation in GRM3 affects cognition, prefrontal glutamate, and risk for schizophrenia
Publisher: Proceedings of the National Academy of Sciences
Date: 13-08-2004
Abstract: GRM3 , a metabotropic glutamate receptor-modulating synaptic glutamate, is a promising schizophrenia candidate gene. In a family-based association study, a common GRM3 haplotype was strongly associated with schizophrenia ( P = 0.0001). Within this haplotype, the A allele of single-nucleotide polymorphism (SNP) 4 (hCV11245618) in intron 2 was slightly overtransmitted to probands ( P = 0.02). We studied the effects of this SNP on neurobiological traits related to risk for schizophrenia and glutamate neurotransmission. The SNP4 A allele was associated with poorer performance on several cognitive tests of prefrontal and hippoc al function. The physiological basis of this effect was assessed with functional MRI, which showed relatively deleterious activation patterns in both cortical regions in control subjects homozygous for the SNP4 A allele. We next looked at SNP4's effects on two indirect measures of prefrontal glutamate neurotransmission. Prefrontal N -acetylaspartate, an in vivo MRI measure related to synaptic activity and closely correlated with tissue glutamate, was lower in SNP4 AA homozygotes. In postmortem human prefrontal cortex, AA homozygotes had lower mRNA levels of the glial glutamate transporter EAAT2, a protein regulated by GRM3 that critically modulates synaptic glutamate. Effects of SNP4 on prefrontal GRM3 mRNA and protein levels were marginal. Resequencing revealed no missense or splice-site SNPs, suggesting that the intronic SNP4 or related haplotypes may exert subtle regulatory effects on GRM3 transcription. These convergent data point to a specific molecular pathway by which GRM3 genotype alters glutamate neurotransmission, prefrontal and hippoc al physiology and cognition, and thereby increased risk for schizophrenia.
Genomic alterations in cultured human embryonic stem cells.
Publisher: Springer Science and Business Media LLC
Date: 04-09-2005
DOI: 10.1038/NG1631
Abstract: Cultured human embryonic stem cell (hESC) lines are an invaluable resource because they provide a uniform and stable genetic system for functional analyses and therapeutic applications. Nevertheless, these iding cells, like other cells, probably undergo spontaneous mutation at a rate of 10(-9) per nucleotide. Because each mutant has only a few progeny, the overall biological properties of the cell culture are not altered unless a mutation provides a survival or growth advantage. Clonal evolution that leads to emergence of a dominant mutant genotype may potentially affect cellular phenotype as well. We assessed the genomic fidelity of paired early- and late-passage hESC lines in the course of tissue culture. Relative to early-passage lines, eight of nine late-passage hESC lines had one or more genomic alterations commonly observed in human cancers, including aberrations in copy number (45%), mitochondrial DNA sequence (22%) and gene promoter methylation (90%), although the latter was essentially restricted to 2 of 14 promoters examined. The observation that hESC lines maintained in vitro develop genetic and epigenetic alterations implies that periodic monitoring of these lines will be required before they are used in in vivo applications and that some late-passage hESC lines may be unusable for therapeutic purposes.
The potential of induced pluripotent stem cells in models of neurological disorders: Implications on future therapy
Publisher: Informa UK Limited
Date: 09-02-2015
DOI: 10.1586/14737175.2015.1013096
Abstract: There is an urgent need for new and advanced approaches to modeling the pathological mechanisms of complex human neurological disorders. This is underscored by the decline in pharmaceutical research and development efficiency resulting in a relative decrease in new drug launches in the last several decades. Induced pluripotent stem cells represent a new tool to overcome many of the shortcomings of conventional methods, enabling live human neural cell modeling of complex conditions relating to aberrant neurodevelopment, such as schizophrenia, epilepsy and autism as well as age-associated neurodegeneration. This review considers the current status of induced pluripotent stem cell-based modeling of neurological disorders, canvassing proven and putative advantages, current constraints, and future prospects of next-generation culture systems for biomedical research and translation.
Functional 3D neural mini-tissues from printed gel-based bioink and human neural stem cells
Publisher: Wiley
Date: 29-03-2016
Abstract: Direct‐write printing of stem cells within biomaterials presents an opportunity to engineer tissue for in vitro modeling and regenerative medicine. Here, a first ex le of constructing neural tissue by printing human neural stem cells that are differentiated in situ to functional neurons and supporting neuroglia is reported. The supporting biomaterial incorporates a novel clinically relevant polysaccharide‐based bioink comprising alginate, carboxymethyl‐chitosan, and agarose. The printed bioink rapidly gels by stable cross‐linking to form a porous 3D scaffold encapsulating stem cells for in situ expansion and differentiation. Differentiated neurons form synaptic contacts, establish networks, are spontaneously active, show a bicuculline‐induced increased calcium response, and are predominantly gamma‐aminobutyric acid expressing. The 3D tissues will facilitate investigation of human neural development, function, and disease, and may be adaptable for engineering other 3D tissues from different stem cell types.
Engineering Human Neural Tissue by 3D Bioprinting
Publisher: Springer New York
Date: 2018
DOI: 10.1007/978-1-4939-7741-3_10
Abstract: Bioprinting provides an opportunity to produce three-dimensional (3D) tissues for biomedical research and translational drug discovery, toxicology, and tissue replacement. Here we describe a method for fabricating human neural tissue by 3D printing human neural stem cells with a bioink, and subsequent gelation of the bioink for cell encapsulation, support, and differentiation to functional neurons and supporting neuroglia. The bioink uniquely comprises the polysaccharides alginate, water-soluble carboxymethyl-chitosan, and agarose. Importantly, the method could be adapted to fabricate neural and nonneural tissues from other cell types, with the potential to be applied for both research and clinical product development.
Human embryonic stem cells have a unique epigenetic signature
Publisher: Cold Spring Harbor Laboratory
Date: 09-08-2006
DOI: 10.1101/GR.5319906
Abstract: Human embryonic stem (hES) cells originate during an embryonic period of active epigenetic remodeling. DNA methylation patterns are likely to be critical for their self-renewal and pluripotence. We compared the DNA methylation status of 1536 CpG sites (from 371 genes) in 14 independently isolated hES cell lines with five other cell types: 24 cancer cell lines, four adult stem cell populations, four lymphoblastoid cell lines, five normal human tissues, and an embryonal carcinoma cell line. We found that the DNA methylation profile clearly distinguished the hES cells from all of the other cell types. A subset of 49 CpG sites from 40 genes contributed most to the differences among cell types. Another set of 25 sites from 23 genes distinguished hES cells from normal differentiated cells and can be used as biomarkers to monitor differentiation. Our results indicate that hES cells have a unique epigenetic signature that may contribute to their developmental potential.
A Call for Standardized Naming and Reporting of Human ESC and iPSC Lines
Publisher: Elsevier BV
Date: 04-2011
DOI: 10.1016/J.STEM.2011.03.002
Abstract: Human embryonic and induced pluripotent stem cell lines are being generated at a rapid pace and now number in the thousands. We propose a standard nomenclature and suggest the use of a centralized database for all cell line names and a minimum set of information for reporting new derivations.
Stem Cell Bioprinting: Functional 3D Neural Mini-Tissues from Printed Gel-Based Bioink and Human Neural Stem Cells (Adv. Healthcare Mater. 12/2016)
Publisher: Wiley
Date: 06-2016
The Generation of Six Clinical-Grade Human Embryonic Stem Cell Lines
Publisher: Elsevier BV
Date: 11-2007
Biomimetic corneal stroma using electro-compacted collagen
Publisher: Elsevier BV
Date: 09-2020
Current and future perspectives on biomaterials for segmental mandibular defect repair
Publisher: Informa UK Limited
Date: 28-03-2023
The binding of [3H]AF-DX 384 is reduced in the caudate-putamen of subjects with schizophrenia
Publisher: Elsevier BV
Date: 04-1999
DOI: 10.1016/S0024-3205(99)00114-9
Abstract: Clinical studies of cholinergic pharmacotherapy, together with the putative role of the muscarinic receptor system in the neurophysiology of human behavior, support a possible muscarinic cholinergic involvement in schizophrenia. The present study has measured the density of [3H]AF-DX 384 labelled receptors (muscarinic M2 and M4) in the caudate-putamen, obtained at autopsy, from 19 subjects who had schizophrenia, and 20 subjects who did not have schizophrenia. [3H]AF-DX 384 binding was reduced in caudate-putamen from schizophrenic subjects (104 +/- 10.3 vs 145 +/- 901 fmol mg(-1) TE mean +/- s.e. p = 0.007). Preliminary analysis of patient drug data as well as rat studies suggest that the reduced [3H]AF-DX 384 binding in caudate-putamen of schizophrenic subjects is not wholly due to antipsychotic drug treatment, or anticholinergic medication for the treatment of extrapyramidal effects. These data suggest that the muscarinic cholinergic system may be involved in the pathology of schizophrenia.
Electrical stimulation-induced osteogenesis of human adipose derived stem cells using a conductive graphene-cellulose scaffold
Publisher: Elsevier BV
Date: 02-2020
DOI: 10.1016/J.MSEC.2019.110312
Abstract: The versatile properties of graphene-based materials are enabling various tissue regeneration, towards meeting an ever increasing demand for replacement tissues due to injury through trauma and disease. In particular, an innate ability for graphene to promote osteogenic differentiation of stem cells, combined with the potential to enhance the biological activity of cells through electrical stimulation (ES) using graphene, supports its use for osteoinduction or reconstruction. In this paper, we describe a miniaturized graphene-cellulose (G-C) scaffold-based device that incorporates electroactive G-C 'paper' within a polystyrene chamber for concomitant cell culture and ES. The G-C electrodes possessed lower impedance and higher charge injection capacity than gold (Au) electrodes, with high stability. By coupling ES with previously reported properties of the G-C scaffolds, we have advanced the platform for improved adipose derived stem cell (ADSC) support and osteogenic differentiation. We anticipate using the G-C scaffold-based ES device for in vitro modelling of osteogenic induction, bone tissue engineering and in vivo bone regeneration towards new therapeutic strategies for bone injury and disease. Furthermore, the device could reasonably be used for ES and culture of other cell types and engineering other tissues.
Standard Culture of Human Embryonic Stem Cells
Publisher: Wiley
Date: 08-06-2007
Electro-mechano responsive properties of gelatin methacrylate (GelMA) hydrogel on conducting polymer electrodes quantified using atomic force microscopy
Publisher: Royal Society of Chemistry (RSC)
Date: 2017
DOI: 10.1039/C7SM00335H
Abstract: Electrical stimulation of hydrogels has been performed to enable micro-actuation or controlled movement of ions and biomolecules such as in drug release applications.
Cholinergic systems and schizophrenia: primary pathology or epiphenomena?
Publisher: Elsevier BV
Date: 07-2001
DOI: 10.1016/S0891-0618(01)00101-6
Abstract: Post mortem schizophrenia research has been driven first by the dopamine and then the glutamate hypotheses. These hypotheses posit primary pathology in pathways dependent upon dopamine or glutamate neurotransmission. Although the dopamine and glutamate hypotheses retain considerable theoretical strength, neurobiological findings of altered dopamine or glutamate activity in schizophrenia do not explain all features of this disorder. A more synthetic approach would suggest that focal pathological change in either the prefrontal cortex or mesial temporal lobe leads to neurochemical changes in multiple neurotransmitter systems. Despite the limited experimental evidence for abnormal cholinergic neurotransmission in psychiatric disorders, increased understanding of the role of acetylcholine in the human brain and its relationship to other neurotransmitter systems has led to a rapidly growing interest in the cholinergic system in schizophrenia. This review focuses on the basic anatomy of the mammalian cholinergic system, and its possible involvement in the neurobiology of schizophrenia. Summaries of cholinergic cell groups, projection pathways, and receptor systems, in the primate and human brain, are followed by a brief discussion of the functional correlations between aberrant cholinergic neurotransmission and the signs and symptoms of schizophrenia.
Engineering in vitro human neural tissue analogs by 3D bioprinting and electrostimulation
Publisher: AIP Publishing
Date: 02-04-2021
DOI: 10.1063/5.0032196
Abstract: There is a fundamental need for clinically relevant, reproducible, and standardized in vitro human neural tissue models, not least of all to study heterogenic and complex human-specific neurological (such as neuropsychiatric) disorders. Construction of three-dimensional (3D) bioprinted neural tissues from native human-derived stem cells (e.g., neural stem cells) and human pluripotent stem cells (e.g., induced pluripotent) in particular is appreciably impacting research and conceivably clinical translation. Given the ability to artificially and favorably regulate a cell's survival and behavior by manipulating its biophysical environment, careful consideration of the printing technique, supporting biomaterial and specific exogenously delivered stimuli, is both required and advantageous. By doing so, there exists an opportunity, more than ever before, to engineer advanced and precise tissue analogs that closely recapitulate the morphological and functional elements of natural tissues (healthy or diseased). Importantly, the application of electrical stimulation as a method of enhancing printed tissue development in vitro, including neuritogenesis, synaptogenesis, and cellular maturation, has the added advantage of modeling both traditional and new stimulation platforms, toward improved understanding of efficacy and innovative electroceutical development and application.
3D Printing of Cytocompatible Graphene/Alginate Scaffolds for Mimetic Tissue Constructs
Publisher: Frontiers Media SA
Date: 17-07-2020
Bioprinting 3D Human Induced Pluripotent Stem Cell Constructs for Multilineage Tissue Engineering and Modeling
Publisher: Springer US
Date: 2020
DOI: 10.1007/978-1-0716-0520-2_17
Abstract: Bioprinting human pluripotent stem cells (PSCs) provides an opportunity to produce three-dimensional (3D) cell-laden constructs with the potential to be differentiated in vitro to all tissue types of the human body. Here, we detail a previously published method for 3D printing human induced pluripotent stem cells (iPSCs also applicable to human embryonic stem cells) within a clinically amenable bioink (also described in Chapter 10 ) that is cross-linked to a 3D construct. The printed iPSCs continue to have self-replicating and multilineage cell induction potential in situ, and the constructs are robust and amenable to different differentiation protocols for fabricating erse tissue types, with the potential to be applied for both research- and clinical-product development.
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