Capture of Neuroepithelial-Like Stem Cells from Pluripotent Stem Cells Provides a Versatile System for In Vitro Production of Human Neurons

Department of Biochemistry, Wellcome Trust Centre for Stem Cell Research, University of Cambridge, Cambridge, United Kingdom.
PLoS ONE (Impact Factor: 3.23). 01/2012; 7(1):e29597. DOI: 10.1371/journal.pone.0029597
Source: PubMed


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.

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Available from: Philipp Koch, Oct 08, 2015
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    • "Combinatorial cell culture is a powerful empirical ‘search engine’ that can be used to discover completely new methods for stem cell differentiation, as well as to optimize or further develop existing protocols, for example by varying the input cell type (e.g. iPS [52], [53] cells, adult stem cells, stable mesenchymal [54], [55] or neuroepithelial [56], [57] intermediates, or differentiated cultures derived by previously determined protocols comprising growth factors or small molecules [19], [58]), the cell culture media components or their concentration, the timing of media changes and many other factors important in cell culture. It can be used to devise protocols that are faster, more productive or cost-effective; protocols that generate higher quality cells, eliminate undefined components such as serum or substitute growth factors with small molecules. "
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    ABSTRACT: We have developed a rapid, bead-based combinatorial screening method to determine optimal combinations of variables that direct stem cell differentiation to produce known or novel cell types having pre-determined characteristics. Here we describe three experiments comprising stepwise exposure of mouse or human embryonic cells to 10,000 combinations of serum-free differentiation media, through which we discovered multiple novel, efficient and robust protocols to generate a number of specific hematopoietic and neural lineages. We further demonstrate that the technology can be used to optimize existing protocols in order to substitute costly growth factors with bioactive small molecules and/or increase cell yield, and to identify in vitro conditions for the production of rare developmental intermediates such as an embryonic lymphoid progenitor cell that has not previously been reported.
    PLoS ONE 09/2014; 9(9):e104301. DOI:10.1371/journal.pone.0104301 · 3.23 Impact Factor
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    • "It is now possible to investigate developmental processes of the human central nervous system (CNS) and the pathogenesis of neural disorders using human PSCs [3] [4] [5] [6]. iPSC-derived NPCs (iPSC-NPCs) can produce both neuronal and glial cells; however, they tend to remain neurogenic for a prolonged period [7] [8]. Extensive research has led to methods for promoting iPSC differentiation toward specific lineages. "
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    ABSTRACT: Here we established a unique human glial cell line, GDC90, derived from a human glioma and demonstrated its utility as a glial scaffold for the polarization and differentiation of human induced pluripotent stem cell-derived neural progenitor cells (iPSC-NPCs). When co-cultured with GDC90 cells, iPSC-NPCs underwent rapid polarization and neurite extension along the radially spreading processes of the GDC90 cells, and showed migratory behavior. This method is potentially useful for detailed examination of neurites or for controlling neurites behavior for regenerative medicine.
    Biochemical and Biophysical Research Communications 04/2014; 447(4). DOI:10.1016/j.bbrc.2014.04.070 · 2.30 Impact Factor
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    • "n , we hope to identify proteins that are only present in a clinical / subclinical neurodevelopmental disorder pheno - type ( see Hölttä et al . , 2012 ; Kroksveen et al . , 2011 ) . Induced pluripotent stem ( iPS ) cells and iPS - derived NES cells from patients are ideal building blocks for the development of disease - specific cellular models ( Falk et al . , 2012 ) . Cellular models built on patient - specific iPS cells have successfully shown disease phenotypes , also when modeling psychiatric disorders ( Christian et al . , 2012 ; Park et al . , 2008 ) . The few reprogramming studies of cells from patients with psychiatric disorders have had encouraging results , proving the principle , as wel"
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    ABSTRACT: Neurodevelopmental disorders affect a substantial minority of the general population. Their origins are still largely unknown, but a complex interplay of genetic and environmental factors causing disturbances of the central nervous system's maturation and a variety of higher cognitive skills is presumed. Only limited research of rather small sample size and narrow scope has been conducted in neurodevelopmental disorders using a twin-differences design. The Roots of Autism and ADHD Twin Study in Sweden (RATSS) is an ongoing project targeting monozygotic twins discordant for categorical or dimensional autistic and inattentive/hyperactive-impulsive phenotypes as well as other neurodevelopmental disorders, and typically developing twin controls. Included pairs are 9 years of age or older, and comprehensively assessed for psychopathology, medical history, neuropsychology, and dysmorphology, as well as structural, functional, and molecular brain imaging. Specimens are collected for induced pluripotent (iPS) and neuroepithelial stem cells, genetic, gut bacteria, protein-/monoamine, and electron microscopy analyses. RATSS's objective is to generate a launch pad for novel surveys to understand the complexity of genotype-environment-phenotype interactions in autism spectrum disorder and attention-deficit hyperactivity disorder (ADHD). By October 2013, RATSS had collected data from 55 twin pairs, among them 10 monozygotic pairs discordant for autism spectrum disorder, seven for ADHD, and four for other neurodevelopmental disorders. This article describes the design, recruitment, data collection, measures, collected pairs' characteristics, as well as ongoing and planned analyses in RATSS. Potential gains of the study comprise the identification of environmentally mediated biomarkers, the emergence of candidates for drug development, translational modeling, and new leads for prevention of incapacitating outcomes.
    Twin Research and Human Genetics 04/2014; 17(3):1-13. DOI:10.1017/thg.2014.12 · 2.30 Impact Factor
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