Unknown,Transcriptomics,Genomics,Proteomics

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A distinctive DNA damage response in human hematopoietic stem cells reveals an apoptosis independent role for p53 in self-renewal

ABSTRACT: Analysis of differentially expressed genes in human Cord Blood cells cultured for 4 days and sorted into CD34+CD90+ and CD34+CD90- fractions. The objective for the experiment was to identify genes related to p53 and DNA damage response pathways Total RNA was isolated from at least 10000 cells after 4 days of culture and sorting

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Milyavsky

PROVIDER: E-GEOD-21830 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Pluripotent mouse embryonic stem (ES) cells multiply in simple monoculture by symmetrical divisions. In vivo, however, stem cells are generally thought to depend on specialised cellular microenvironments and to undergo predominantly asymmetric divisions. Ex vivo expansion of pure populations of tissue stem cells has proven elusive. Neural progenitor cells are propagated in combination with differentiating progeny in floating clusters called neurospheres. The proportion of stem cells in neurospheres is low, however, and they cannot be directly observed or interrogated. Here we demonstrate that the complex neurosphere environment is dispensable for stem cell maintenance, and that the combination of fibroblast growth factor 2 (FGF-2) and epidermal growth factor (EGF) is sufficient for derivation and continuous expansion by symmetrical division of pure cultures of neural stem (NS) cells. NS cells were derived first from mouse ES cells. Neural lineage induction was followed by growth factor addition in basal culture media. In the presence of only EGF and FGF-2, resulting NS cells proliferate continuously, are diploid, and clonogenic. After prolonged expansion, they remain able to differentiate efficiently into neurons and astrocytes in vitro and upon transplantation into the adult brain. Colonies generated from single NS cells all produce neurons upon growth factor withdrawal. NS cells uniformly express morphological, cell biological, and molecular features of radial glia, developmental precursors of neurons and glia. Consistent with this profile, adherent NS cell lines can readily be established from foetal mouse brain. Similar NS cells can be generated from human ES cells and human foetal brain. The extrinsic factors EGF plus FGF-2 are sufficient to sustain pure symmetrical self-renewing divisions of NS cells. The resultant cultures constitute the first known example of tissue-specific stem cells that can be propagated without accompanying differentiation. These homogenous cultures will enable delineation of molecular mechanisms that define a tissue-specific stem cell and allow direct comparison with pluripotent ES cells.

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A distinctive DNA damage response in human hematopoietic stem cells reveals an apoptosis independent role for p53 in self-renewal

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