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MRNA sequencing of clinical-grade neural stem cells derived from human ES cells

ABSTRACT: mRNA sequencing of clinical-grade neural stem cells derived from human ES cells

PROVIDER: PRJNA559188 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9922680_1.fastq.gz	Fastqsanger.gz
	SRR9922680_2.fastq.gz	Fastqsanger.gz
	SRR9922671_1.fastq.gz	Fastqsanger.gz
	SRR9922671_2.fastq.gz	Fastqsanger.gz
	SRR9922681.fastq.gz	Fastqsanger.gz

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Project description:Human embryonic stem cells not only provide a continuous cell source for potential cell therapy but also offer a system to unveil events of embryonic development in humans. This proposal will examine how the earliest neural cells, neuroepithelia, are specified from the naïve ES cells, and test the hypothesis that neural specification in humans employs a similar mechanism as in other vertebrates. We will first re-create in culture the developmental events of the first 2-3 weeks of human embryonic development during which ES cells will be differentiated through the stages of embryoid bodies, primitive ectoderm cells, neural tube-like rosette cells. The stage-specific events will be defined by DNA microarray analysis along with the characteristic morphologic changes. This study will lead to an optimized procedure for generating enriched neural precursor cells, which will lay the groundwork for potential use of human ES cells in the treatment of neurological injuries and diseases. Aim 1: Establish a stepwise neural differentiation system from human ES cells. Aim 2. Determine the mechanism of Neural Specification in humans. Aim 3: Define the identity and function of ES cell-derived neural precursor cells. ES cells offer an alternative approach to study early developmental events in humans. This however requires re-creating the neural differentiation process in culture. Cell fate specification is determined by interactions between environmental factors and intrinsic signals. Our preliminary data suggest that the temporal pattern of neural differentiation and, to some degree, the spatial organization of neural precursors from human ES cells recapitulate in vivo neural development. We hypothesize that the intrinsic neural specification program may be preserved in culture, which offers a controlled system to examine the effect of extrinsic signals on neural specification. Distinctive morphological changes along the neural differentiation pathway are presumably accompanied by molecular changes. DNA microarray analysis will be used to determine the gene expression pattern by cells at each of the given stages. By analogy with early embryonic development and using morphological and antigenic markers, we can now subdivide the human ES cell neural differentiation process into four identifiable stages: ES, EB, PEL cells, and neural rosette cells. This definition is based on the assumption that early human development is the same as in other species, and employs the limited known markers from mouse ES cells. We will systematically investigate the molecular profile of cells at each of the neural differentiation stages using DNA microarray analysis. Total RNAs will be extracted from cells at the following developmental stages: ES cells, EBs grown in suspension (d6), PEL (d10) stage and neural rosettes (d17). Since the neural rosette culture contains non-neural lineage cells, we will separate the neural rosette cells from the surrounding non-neural cells through differential enzymatic response and differential adhesion. Three independent biological replicates consisting of three pooled experiments will be run for each of the four developmental time points, for a total of twelve chips. Keywords: time-course

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MRNA sequencing of clinical-grade neural stem cells derived from human ES cells

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