Austin smith biologist

His work has laid the foundations for the development of potential stem cell therapies for degenerative diseases in humans. Having identified the intrinsic and extrinsic factors that support pluripotency, Austin developed a culture medium for mouse stem cells that allows them to renew themselves indefinitely.

Austin has extended his work into the more challenging area of human embryonic stem cells hESCs and induced pluripotent stem cells iPSCs. He coordinates Europe-wide programmes of stem cell research, and in January he was awarded the prestigious Louis-Jeantet Prize for Medicine.

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Article Navigation. This site. Google Scholar. Marc K. Saba-El-Leil , Marc K. Marwa Almousailleakh , Marwa Almousailleakh. Jason Wray , Jason Wray. Sylvain Meloche , Sylvain Meloche. Author and article information. Marwa Almousailleakh. Jason Wray. Sylvain Meloche. Accepted: 12 Jun Online Issn: Development 16 : — Article history Accepted:. Cite Icon Cite. Table 1. Primers and conditions for reverse transcriptase PCR.

Forward primer. Reverse primer. Product bp. Annealing temp. View Large. View large Download slide. Brook, F. Burdon, T. Chambers, I. Chen, Y. Cheng, A.

Corson, L. Davies, S. Evans, M. Goto, H. Hamazaki, T. Haub, O. Ivanova, N. Jirmanova, L. Keller, G. Kouskoff, V. Kretzschmar, M. Kuroda, H. Lowell, S. Ma, Y. Martin, G. Mohammadi, M. Nishikawa, S. Niwa, H. Pera, E. Qu, C. Rathjen, J. Rogers, M. Roux, P. Saba-El-Leil, M. Shen, M. Smith, A.

Stavridis, M. Stern, C. Streit, A. Consequently, they harbour latent potential, as manifested in teratocarcinogenesis or direct conversion into pluripotent stem cells in vitro.

Thus, we suggest that there exists an unbroken cycle of pluripotency, naive in the early epiblast and latent in the germline, that is sustained by a shared transcription factor network.

Widespread resetting of DNA methylation in glioblastoma-initiating cells suppresses malignant cellular behavior in a lineage-dependent manner. Genes Dev , 27 , Abstract: Widespread resetting of DNA methylation in glioblastoma-initiating cells suppresses malignant cellular behavior in a lineage-dependent manner. Epigenetic changes are frequently observed in cancer.

However, their role in establishing or sustaining the malignant state has been difficult to determine due to the lack of experimental tools that enable resetting of epigenetic abnormalities. To address this, we applied induced pluripotent stem cell iPSC reprogramming techniques to invoke widespread epigenetic resetting of glioblastoma GBM -derived neural stem GNS cells.

Surprisingly, despite such global epigenetic reconfiguration, GiPSC-derived neural progenitors remained highly malignant upon xenotransplantation. Only when GiPSCs were directed to nonneural cell types did we observe sustained expression of reactivated tumor suppressors and reduced infiltrative behavior. These data suggest that imposing an epigenome associated with an alternative developmental lineage can suppress malignant behavior. However, in the context of the neural lineage, widespread resetting of GBM-associated epigenetic abnormalities is not sufficient to override the cancer genome.

PLoS Genet , 8 , ee Embryonic stem cells and induced pluripotent stem cells represent potentially important therapeutic agents in regenerative medicine. Complex interlinked transcriptional and signaling networks control the fate of these cells towards maintenance of pluripotency or differentiation.

In this study we have focused on how mouse embryonic stem cells begin to differentiate and lose pluripotency and, in particular, the role that the ERK MAP kinase and GSK3 signaling pathways play in this process. Through a genome-wide siRNA screen we have identified more than genes involved in loss of pluripotency and promoting the onset of differentiation. More detailed analysis identified MAP kinase phosphatases as a focal point of regulation and demonstrated an important role for these enzymes in controlling ERK activation kinetics and subsequently determining early embryonic stem cell fate decisions.

Capture of neuroepithelial-like stem cells from pluripotent stem cells provides a versatile system for in vitro production of human neurons. PLoS One , 7 , ee Abstract: Capture of neuroepithelial-like stem cells from pluripotent stem cells provides a versatile system for in vitro production of human neurons. Human embryonic stem cells hESC and induced pluripotent stem cells iPSC provide new prospects for studying human neurodevelopment and modeling neurological disease.

In particular, iPSC-derived neural cells permit a direct comparison of disease-relevant molecular pathways in neurons and glia derived from patients and healthy individuals. A prerequisite for such comparative studies are robust protocols that efficiently yield standardized populations of neural cell types. Here we show that long-term self-renewing neuroepithelial-like stem cells lt-NES cells derived from 3 hESC and 6 iPSC lines in two independent laboratories exhibit consistent characteristics including i continuous expandability in the presence of FGF2 and EGF; ii stable neuronal and glial differentiation competence; iii characteristic transcription factor profile; iv hindbrain specification amenable to regional patterning; v capacity to generate functionally mature human neurons.

We further show that lt-NES cells are developmentally distinct from fetal tissue-derived radial glia-like stem cells. We propose that lt-NES cells provide an interesting tool for studying human neurodevelopment and may serve as a standard system to facilitate comparative analyses of hESC and hiPSC-derived neural cells from control and diseased genetic backgrounds.

Culture parameters for stable expansion, genetic modification and germline transmission of rat pluripotent stem cells. Biol Open , 1 , Abstract: Culture parameters for stable expansion, genetic modification and germline transmission of rat pluripotent stem cells.

The ability of cultured pluripotent cells to contribute to the germline of chimaeric animals is essential to their utility for genetic manipulation. In the three years since rat embryonic stem ES cells were first reported the anticipated proliferation of genetically modified rat models from this new resource has not been realised. Culture instability, karyotypic anomalies, and strain variation are postulated to contribute to poor germline colonisation capacity.

The resolution of these issues is essential to bring pluripotent cell-based genetic manipulation technology in the rat to the level of efficiency achieved in the mouse. Recent reports have described various alternative methods to maintain rat ES cells that include provision of additional small molecules and selective passaging methods. We demonstrate genetic modification, clonal expansion and transmission through the germline of rat ES and EG cell lines.

We also describe a marked preference for full-term chimaera contribution when SD strain blastocysts are used as recipients for either DA or SD pluripotent stem cells. Cell Stem Cell , 11 4 , Inhibition of glycogen synthase kinase-3 Gsk3 supports mouse embryonic stem cells ESCs by modulating Tcf3, but the critical targets downstream of Tcf3 are unclear. We analyzed the intersection between genome localization and transcriptome data sets to identify genes repressed by Tcf3.

Among these, manipulations of Esrrb gave distinctive phenotypes in functional assays. Knockdown and knockout eliminated response to Gsk3 inhibition, causing extinction of pluripotency markers and loss of colony forming capability.

Conversely, forced expression phenocopied Gsk3 inhibition or Tcf3 deletion by suppressing differentiation and sustaining self-renewal. Thus the nuclear receptor Esrrb is necessary and sufficient to mediate self-renewal downstream of Gsk3 inhibition.

These findings highlight a key role for Esrrb in regulating the naive pluripotent state and illustrate compensation among the core pluripotency factors. Germline potential of parthenogenetic haploid mouse embryonic stem cells. Abstract: Germline potential of parthenogenetic haploid mouse embryonic stem cells. Haploid embryonic stem cells ESCs have recently been derived from parthenogenetic mouse embryos and offer new possibilities for genetic screens.

The ability of haploid ESCs to give rise to a wide range of differentiated cell types in the embryo and in vitro has been demonstrated. However, it has remained unclear whether haploid ESCs can contribute to the germline.

Here, we show that parthenogenetic haploid ESCs at high passage have robust germline competence enabling the production of transgenic mouse strains from genetically modified haploid ESCs.

We also show that differentiation of haploid ESCs in the embryo correlates with the gain of a diploid karyotype and that diploidisation is the result of endoreduplication and not cell fusion. By contrast, we find that a haploid karyotype is maintained when differentiation to an extra-embryonic fate is forced by induction of Gata6. Human hypoblast formation is not dependent on FGF signalling.

Dev Biol , , Abstract: Human hypoblast formation is not dependent on FGF signalling. Mouse embryos segregate three different lineages during preimplantation development: trophoblast, epiblast and hypoblast. These differentiation processes are associated with restricted expression of key transcription factors Cdx2, Oct4, Nanog and Gata6. The mechanisms of segregation have been extensively studied in the mouse, but are not as well characterised in other species.

In the human embryo, hypoblast differentiation has not previously been characterised. Here we demonstrate co-exclusive immunolocalisation of Nanog and Gata4 in human blastocysts, implying segregation of epiblast and hypoblast, as in rodent embryos.

However, the formation of hypoblast in the human is apparently not dependent upon FGF signalling, in contrast to rodent embryos. Imaging, quantification and visualization of spatio-temporal patterning in mESC colonies under different culture conditions. Bioinformatics , 28 , ii Abstract: Imaging, quantification and visualization of spatio-temporal patterning in mESC colonies under different culture conditions.

It is well recognized that different, established protocols for the maintenance of mESC pluripotency support morphologically and functionally different cell cultures. However, it is unclear how characteristic properties of cell colonies develop over time and how they are re-established after cell passage depending on the culture conditions.

Furthermore, it appears that cell colonies have an internal structure with respect to cell size, marker expression or biomechanical properties, which is not sufficiently understood. The analysis of these phenotypic properties is essential for a comprehensive understanding of mESC development and ultimately requires a bioinformatics approach to guarantee reproducibility and high-throughput data analysis. RESULTS: We developed an automated image analysis and colony tracking framework to obtain an objective and reproducible quantification of structural properties of cell colonies as they evolve in space and time.

In particular, we established a method that quantifies changes in colony shape and internal motion using fluid image registration and image segmentation. The methodology also allows to robustly track motion, splitting and merging of colonies over a sequence of images. Our results provide a first quantitative assessment of temporal mESC colony formation and estimates of structural differences between colony growth under different culture conditions.

Furthermore, we provide a stream-based visualization of structural features of individual colonies over time for the whole experiment, facilitating visual comprehension of differences between experimental conditions. Thus, the presented method establishes the basis for the model-based analysis of mESC colony development.

It can be easily extended to integrate further functional information using fluorescence signals and differentiation markers. The tool is freely available from the authors. We will also provide the source code upon request. Induction of superficial cortical layer neurons from mouse embryonic stem cells by valproic acid.

Neurosci Res , 72 , Abstract: Induction of superficial cortical layer neurons from mouse embryonic stem cells by valproic acid. Within the developing mammalian cortex, neural progenitors first generate deep-layer neurons and subsequently more superficial-layer neurons, in an inside-out manner.

It has been reported recently that mouse embryonic stem cells mESCs can, to some extent, recapitulate cortical development in vitro, with the sequential appearance of neurogenesis markers resembling that in the developing cortex.

However, mESCs can only recapitulate early corticogenesis; superficial-layer neurons, which are normally produced in later developmental periods in vivo, are under-represented. This failure of mESCs to reproduce later corticogenesis in vitro implies the existence of crucial factor s that are absent or uninduced in existing culture systems. Here we show that mESCs can give rise to superficial-layer neurons efficiently when treated with valproic acid VPA , a histone deacetylase inhibitor.

VPA treatment increased the production of Cux1-positive superficial-layer neurons, and decreased that of Ctip2-positive deep-layer neurons.

These results shed new light on the mechanisms of later corticogenesis. Inhibition of glycogen synthase kinase-3 alleviates Tcf3 repression of the pluripotency network and increases embryonic stem cell resistance to differentiation. Nat Cell Biol , 14 , Nat Commun , 3 , Induced pluripotency depends on cooperativity between expression of defined factors. The latter also determines the pluripotent cell state. Here we show that sufficient activation of. Moreover, even in the presence of FGF.

Nichols J, Smith A Pluripotency in the embryo and in culture. Cold Spring Harb Perspect Biol , 4 , aa Abstract: Pluripotency in the embryo and in culture.

Specific cells within the early mammalian embryo have the capacity to generate all somatic lineages plus the germline. This property of pluripotency is confined to the epiblast, a transient tissue that persists for only a few days. In vitro, however, pluripotency can be maintained indefinitely through derivation of stem cell lines.

Pluripotent stem cells established from the newly formed epiblast are known as embryonic stem cells ESCs , whereas those generated from later stages are called postimplantation epiblast stem cells EpiSCs.

These different classes of pluripotent stem cell have distinct culture requirements and gene expression programs, likely reflecting the dynamic development of the epiblast in the embryo. In this chapter we review current understanding of how the epiblast forms and relate this to the properties of derivative stem cells. We discuss whether ESCs and EpiSCs are true counterparts of different phases of epiblast development or are culture-generated phenomena.

Self-organizing circuitry and emergent computation in mouse embryonic stem cells. Stem Cell Res , 8 , Abstract: Self-organizing circuitry and emergent computation in mouse embryonic stem cells. Pluripotency is a cellular state of multiple options. Here, we highlight the potential for self-organization to contribute to stem cell fate computation. A new way of considering regulatory circuitry is presented that describes the expression of each transcription factor TF as a branching process that propagates through time, interacting and competing with others.

We explain how this phenomenon provides a valid description of whole genome regulatory circuit dynamics. The hypothesis of exploratory stem cell decision-making proposes that critical-like self-organization also called rapid self-organized criticality provides the backbone for cell fate computation in regulative embryos and pluripotent stem cells.

Unspecific amplification of TF expression is predicted to initiate this self-organizing circuitry, where cascades of gene expression propagate and may interact either synergistically or antagonistically. The emergent and highly dynamic circuitry is affected by various sources of selection pressure, such as the expression of TFs with disproportionate influence over other genes, and extrinsic biological and physical stimuli that differentially modulate particular gene expression cascades.

Extrinsic conditions continuously trigger waves of transcription that ripple throughout regulatory networks on multiple spatiotemporal scales, providing the context within which circuitry self-organizes. Neurobiol Dis , 46 , The establishment of an in vitro model that recapitulates features of the disease may help understanding the cellular and molecular events that trigger disease manifestations.

Here, we describe the generation and characterization of a series of induced pluripotent stem iPS cells derived from patients with HD, including two rare homozygous genotypes and one heterozygous genotype. We used lentiviral technology to transfer key genes for inducing reprogramming. To confirm pluripotency and differentiation of iPS cells, we used PCR amplification and immunocytochemistry to measure the expression of marker genes in embryoid bodies and neurons.

We also analyzed teratomas that formed in iPS cell-injected mice. We found that the length of the pathological CAG repeat did not increase during reprogramming, after long term growth in vitro, and after differentiation into neurons.

In addition, we observed no differences between normal and mutant genotypes in reprogramming, growth rate, caspase activation or neuronal differentiation. However, we observed a significant increase in lysosomal activity in HD-iPS cells compared to control iPS cells, both during self-renewal and in iPS-derived neurons.

In conclusion, we have established stable HD-iPS cell lines that can be used for investigating disease mechanisms that underlie HD. In the future, these cells may provide the basis for a powerful platform for drug screening and target identification in HD. The transcriptional and epigenomic foundations of ground state pluripotency.

Abstract: The transcriptional and epigenomic foundations of ground state pluripotency. Mouse embryonic stem ES cells grown in serum exhibit greater heterogeneity in morphology and expression of pluripotency factors than ES cells cultured in defined medium with inhibitors of two kinases Mek and GSK3 , a condition known as "2i" postulated to establish a naive ground state.

We show that the transcriptome and epigenome profiles of serum- and 2i-grown ES cells are distinct. Nonetheless, serum- and 2i-grown ES cells have similar differentiation potential. Precocious transcription of developmental genes in 2i is restrained by RNA polymerase II promoter-proximal pausing.

These findings suggest that transcriptional potentiation and a permissive chromatin context characterize the ground state and that exit from it may not require a metastable intermediate or multilineage priming.

Treatment of a mouse model of spinal cord injury by transplantation of human induced pluripotent stem cell-derived long-term self-renewing neuroepithelial-like stem cells. Stem Cells , 30 , Abstract: Treatment of a mouse model of spinal cord injury by transplantation of human induced pluripotent stem cell-derived long-term self-renewing neuroepithelial-like stem cells. Because of their ability to self-renew, to differentiate into multiple lineages, and to migrate toward a damaged site, neural stem cells NSCs , which can be derived from various sources such as fetal tissues and embryonic stem cells, are currently considered to be promising components of cell replacement strategies aimed at treating injuries of the central nervous system, including the spinal cord.

Despite their efficiency in promoting functional recovery, these NSCs are not homogeneous and possess variable characteristics depending on their derivation protocols.

The advent of induced pluripotent stem iPS cells has provided new prospects for regenerative medicine. We used a recently developed robust and stable protocol for the generation of long-term, self-renewing, neuroepithelial-like stem cells from human iPS cells hiPS-lt-NES cells , which can provide a homogeneous and well-defined population of NSCs for standardized analysis.

Here, we show that transplanted hiPS-lt-NES cells differentiate into neural lineages in the mouse model of spinal cord injury SCI and promote functional recovery of hind limb motor function. Furthermore, using two different neuronal tracers and ablation of the transplanted cells, we revealed that transplanted hiPS-lt-NES cell-derived neurons, together with the surviving endogenous neurons, contributed to restored motor function. Both types of neurons reconstructed the corticospinal tract by forming synaptic connections and integrating neuronal circuits.

Nat Cell Biol , 13 , Abstract: Inhibition of glycogen synthase kinase-3 alleviates Tcf3 repression of the pluripotency network and increases embryonic stem cell resistance to differentiation. Self-renewal of rodent embryonic stem cells is enhanced by partial inhibition of glycogen synthase kinase-3 Gsk3; refs 1, 2.

Tcf3 localizes to many pluripotency genes in embryonic stem cells. We conclude that Gsk3 inhibition stabilizes the embryonic stem cell state primarily by reducing repressive influence on the core pluripotency network. J Cell Sci , , Mouse and human central nervous system progenitor cells can be propagated extensively ex vivo as stem cell lines.

For the rat, however, in vitro expansion has proven to be problematic owing to proliferation arrest and differentiation. Here, we analyse the establishment, in adherent culture, of undifferentiated tripotent neural stem NS cell lines derived from rat foetal brain and spinal cord. Rat NS cells invariably undergo growth arrest and apparent differentiation after several passages; however, conditioned medium from proliferating cultures can overcome this block, enabling continuous propagation of undifferentiated rat NS cells.

We found that dormancy is induced by autocrine production of bone morphogenetic proteins BMPs. Accordingly, the BMP antagonist noggin can replace conditioned medium to sustain continuous self-renewal.

Noggin can also induce dormant cells to re-enter the cell cycle, upon which they reacquire neurogenic potential. We further show that fibroblast growth factor 2 FGF2 is required to suppress terminal astrocytic differentiation and maintain stem cell potency during dormancy. These findings highlight an extrinsic regulatory network, comprising BMPs, BMP antagonists and FGF2 signals, that governs the proliferation, dormancy and differentiation of rat NS cells and which can be manipulated to enable long-term clonogenic self-renewal.

Nanog overcomes reprogramming barriers and induces pluripotency in minimal conditions. Curr Biol , 21 , Abstract: Nanog overcomes reprogramming barriers and induces pluripotency in minimal conditions.

Induced pluripotency requires the expression of defined factors and culture conditions that support the self-renewal of embryonic stem ES cells. Nanog enables somatic cell reprogramming in serum-free medium supplemented with LIF, a culture condition which does not support induced pluripotency or the self-renewal of ES cells, and is sufficient to reprogram epiblast-derived stem cells to naive pluripotency in serum-free medium alone.

Nanog also enhances reprogramming in cooperation with kinase inhibition or 5-aza-cytidine, a small molecule inhibitor of DNA methylation. These results highlight the capacity of Nanog to overcome multiple barriers to reprogramming and reveal a synergy between Nanog and chemical inhibitors that promote reprogramming.

We conclude that Nanog induces pluripotency in minimal conditions. This provides a strategy for imposing naive pluripotency in mammalian cells independently of species-specific culture requirements. Sox2 and Pax6 maintain the proliferative and developmental potential of gliogenic neural stem cells in vitro. Glia , 59 , Abstract: Sox2 and Pax6 maintain the proliferative and developmental potential of gliogenic neural stem cells in vitro.

Radial-glia-like neural stem NS cells may be derived from neural tissues or via differentiation of pluripotent embryonic stem ES cells. However, the mechanisms controlling NS cell propagation and differentiation are not yet fully understood. Here we investigated the roles of Sox2 and Pax6, transcription factors widely expressed in central nervous system CNS progenitors, in mouse NS cells.

Conditional deletion of either Sox2 or Pax6 in forebrain-derived NS cells reduced their clonogenicity in a gene dosage-dependent manner. Cells heterozygous for either gene displayed moderate proliferative defects, which may relate to human pathologies attributed to SOX2 or PAX6 deficiencies.

In the complete absence of Sox2, cells exited the cell cycle with concomitant downregulation of neural progenitor markers Nestin and Blbp. This occurred despite expression of the close relative Sox3. Ablation of Pax6 also caused major proliferative defects. However, a subpopulation of cells was able to expand continuously without Pax6.

These Pax6-null cells retained progenitor markers but had altered morphology. They exhibited compromised differentiation into astrocytes and oligodendrocytes, highlighting that the role of Pax6 extends beyond neurogenic competence.

Overall these findings indicate that Sox2 and Pax6 are both critical for self-renewal of differentiation-competent radial glia-like NS cells. The liberation of embryonic stem cells. PLoS Genet , 7 , ee Abstract: The liberation of embryonic stem cells.

Mouse embryonic stem ES cells are defined by their capacity to self-renew and their ability to differentiate into all adult tissues including the germ line. Along with efficient clonal propagation, these properties have made them an unparalleled tool for manipulation of the mouse genome. Traditionally, mouse ES mES cells have been isolated and cultured in complex, poorly defined conditions that only permit efficient derivation from the mouse strain; genuine ES cells have not been isolated from another species in these conditions.

Recently, use of small molecule inhibitors of glycogen synthase kinase 3 Gsk3 and the Fgf-MAPK signaling cascade has permitted efficient derivation of ES cells from all tested mouse strains. Subsequently, the first verified ES cells were established from a non-mouse species, Rattus norvegicus. Here, we summarize the advances in our understanding of the signaling pathways regulating mES cell self-renewal that led to the first derivation of rat ES cells and highlight the new opportunities presented for transgenic modeling on diverse genetic backgrounds.

We also comment on the implications of this work for our understanding of pluripotent stem cells across mammalian species. The origin and identity of embryonic stem cells. Abstract: The origin and identity of embryonic stem cells. Embryonic stem ES cells are used extensively in biomedical research and as a model with which to study early mammalian development, but their exact origin has been subject to much debate.

They are routinely derived from pre-implantation embryos, but it has been suggested that the cells that give rise to ES cells might arise from epiblast cells that are already predisposed to a primordial germ cell PGC fate, which then progress to ES cell status via the PGC lineage.

Based on recent findings, we propose here that ES cells can be derived directly from early epiblast cells and that ES cells might arise via two different routes that are dictated by their culture conditions. Nature , , Guo G, Smith A A genome-wide screen in EpiSCs identifies Nr5a nuclear receptors as potent inducers of ground state pluripotency.

Embryonic germ cells from mice and rats exhibit properties consistent with a generic pluripotent ground state. Abstract: Embryonic germ cells from mice and rats exhibit properties consistent with a generic pluripotent ground state.

Pluripotent stem cell lines can also be derived from unipotent primordial germ cells via a poorly understood process of epigenetic reprogramming. These are termed embryonic germ EG cells to denote their distinct origin. Here we investigate whether EG cell self-renewal and derivation are supported by 2i. We report that mouse EG cells can be established with high efficiency using 2i in combination with the cytokine leukaemia inhibitory factor LIF.

Furthermore, addition of fibroblast growth factor or stem cell factor is unnecessary using 2i-LIF. The derived EG cells contribute extensively to healthy chimaeric mice, including to the germline. Using the same conditions, we describe the first derivations of EG cells from the rat. Rat EG cells express a similar marker profile to rat and mouse ES cells. They have a diploid karyotype, can be clonally expanded and genetically manipulated, and are competent for multilineage colonisation of chimaeras.

These findings lend support to the postulate of a conserved molecular ground state in pluripotent rodent cells. Imaging-based chemical screens using normal and glioma-derived neural stem cells.

Biochem Soc Trans , 38 , Abstract: Imaging-based chemical screens using normal and glioma-derived neural stem cells.

The development of optimal culture methods for embryonic, tissue and cancer stem cells is a critical foundation for their application in drug screening. We previously described defined adherent culture conditions that enable expansion of human radial glia-like fetal NS neural stem cells as stable cell lines. Similar protocols proved effective in the establishment of tumour-initiating stem cell lines from the human brain tumour glioblastoma multiforme, which we termed GNS glioma NS cells.

Others have also recently derived more primitive human NS cell lines with greater neuronal subtype differentiation potential than NS cells, which have similarities to the early neuroepithelium, named NES neuroepithelial stem cells.

In the present paper, we discuss the utility of these cells for chemical screening, and describe methods for a simple high-content live-image-based platform. For the GNS cells, we screened a panel of compounds and confirmed our previous finding of a cytotoxic effect of modulators of neurotransmitter signalling pathways.

These studies provide a framework for future higher-throughput screens. Isolation and propagation of enteric neural crest progenitor cells from mouse embryonic stem cells and embryos. Abstract: Isolation and propagation of enteric neural crest progenitor cells from mouse embryonic stem cells and embryos.

Neural crest is a source of diverse cell types, including the peripheral nervous system. The transcription factor Sox10 is expressed throughout early neural crest. We exploited Sox10 reporter and selection markers created by homologous recombination to investigate the generation, maintenance and expansion of neural crest progenitors.

SoxGFP-positive cells are produced transiently from mouse embryonic stem ES cells by treatment with retinoic acid in combination with Fgf8b and the cytokine leukaemia inhibitory factor Lif. They differentiate into peripheral neurons in vitro and are able to colonise the enteric network in organotypic gut cultures.

Neural crest cells purified from embryos using the Sox10 reporter also survive in NWLE, but progressively succumb to differentiation. We therefore applied selection to eliminate differentiating cells. Soxselected cells could be clonally expanded, cryopreserved, and multiplied for over 50 days in adherent culture. They remained neurogenic in vitro and in foetal gut grafts.

Generation of neural crest from mouse ES cells opens a new route to the identification and validation of determination factors. Furthermore, the ability to propagate undifferentiated progenitors creates an opportunity for experimental dissection of the stimuli and molecular circu that govern neural crest lineage progression.

Ectopic expression of defined sets of genetic factors can reprogram somatic cells to create induced pluripotent stem iPS cells. The capacity to direct human iPS cells to specific differentiated lineages and to their progenitor populations can be used for disease modeling, drug discovery, and eventually autologous cell replacement therapies. During mouse cardiogenesis, the major lineages of the mature heart, cardiomyocytes, smooth muscle cells, and endothelial cells arise from a common, multipotent cardiovascular progenitor expressing the transcription factors Isl1 and Nkx2.

These results support the possibility to use patient-specific iPS-generated cardiovascular progenitors as a model to elucidate the pathogenesis of congenital and acquired forms of heart diseases. Bellin, M. Jung, C. Thies, T. Takashima, Y. Bernshausen, A. Schiemann, M. Fischer, S. Moosmang, S. Smith, A. Lam, J. Laugwitz, K. Pluripotent stem cells: private obsession and public expectation. The ground state of pluripotency. Abstract: The ground state of pluripotency. Pluripotency is defined as the capacity of individual cells to initiate all lineages of the mature organism in response to signals from the embryo or cell culture environment.

A pluripotent cell has no predetermined programme; it is a blank slate. This is the foundation of mammalian development and of ES embryonic stem cell biology. How is pluripotency acquired and maintained? Suppressing activation of ERKs extracellular-signal-regulated kinases is critical to establishing and sustaining ES cells.

Inhibition of GSK3 glycogen synthase kinase 3 reinforces this effect. We review the effect of selective kinase inhibitors on pluripotent cells and consider how these effects are mediated. We propose that ES cells represent a ground state, meaning a basal proliferative state that is free of epigenetic restriction and has minimal requirements for extrinsic stimuli.

CD Prominin negative human neural stem cells are clonogenic and tripotent. PLoS One , 4 , ee Abstract: CD Prominin negative human neural stem cells are clonogenic and tripotent. However, the assumption that CD is expressed constitutively in neural precursor cells has not been examined.

After fractionation by flow cytometry, clonogenic tripotent cells are found in populations negative or positive for either marker. A similar pattern is apparent in mouse NS cell lines. Compared to mouse NS cells, however, human NS cell cultures harbour an increased proportion of CD negative cells and display a longer doubling time.

Non-proliferating NS cells remain undifferentiated and at least some of them are capable of re-entry into the cell cycle and subsequent continuous expansion. Design principles of pluripotency. Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens.

Cell Stem Cell , 4 , Abstract: Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Human brain tumors appear to have a hierarchical cellular organization suggestive of a stem cell foundation. In vitro expansion of the putative cancer stem cells as stable cell lines would provide a powerful model system to study their biology.

Here, we demonstrate routine and efficient derivation of adherent cell lines from malignant glioma that display stem cell properties and initiate high-grade gliomas following xenotransplantation.

Significantly, glioma neural stem GNS cell lines from different tumors exhibit divergent gene expression signatures and differentiation behavior that correlate with specific neural progenitor subtypes.