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Cloning and Variation of Ground State Intestinal Stem Cells Super [expression & SNP studies]

ABSTRACT: Cloning and Variation of Ground State Intestinal Stem Cells Super [expression & SNP studies]

PROVIDER: PRJNA277854 | ENA |

REPOSITORIES: ENA

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Cloning and Variation of Ground State Intestinal Stem Cells Super [expression & SNP studies]

Project description:This SuperSeries is composed of the SubSeries listed below.

2015-05-29 | GSE66749 | GEO

Cloning and Variation of Ground State Intestinal Stem Cells

Project description:Despite major advances with embryonic and induced pluripotent stem cells or lineage-committed, p63-expressing stem cells of stratified epithelia, we know less about the indigenous stem cells of the gastrointestinal tract, pancreas, liver, and other columnar epithelia which collectively resist cloning in their elemental states. Here we demonstrate the cloning of highly immature epithelial stem cells from defined regions of the human intestine and colon. We show that single cell-derived pedigrees can be propagated indefinitely while often sustaining minimal copy number and sequence variation. Despite prolonged cultivation, these pedigrees from disparate regions of the intestinal tract respond to identical differentiation signals by formation of epithelia with eponymous structural and gene expression features. These data suggest developmental patterning of cell-autonomous commitment programs in stem cells that enforce specialization along the gastrointestinal tract and predict the utility of these cells in disease modeling and regenerative medicine.

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Project description:Stem cells of the gastrointestinal tract, pancreas, liver, and other columnar epithelia collectively resist cloning in their elemental states. Here we demonstrate the cloning and propagation of highly clonogenic, “ground state” stem cells of the human intestinal and colon. To assess the genomic stability of our intestinal stem cells, we serially passaged five independent intestinal stem cell pedigrees derived from the ileum of one late fetal demise case and tested them after 50 (passage 5; P5) and 250 days (P25) of continuous proliferation. DNA from cells at these passages cells was analyzed by SNP array based copy number variation analysis (CNV). Although at P25, there are aneuploidy in some of pedigrees, there are very few CNV at P5 in all pedigrees.

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Copy number variation of human intestinal stem cells with different passage in culture (two pedigrees, passage 5, 10, 15 and 20)

Project description:Stem cells of the gastrointestinal tract, pancreas, liver, and other columnar epithelia collectively resist cloning in their elemental states. Here we demonstrate the cloning and propagation of highly clonogenic, “ground state” stem cells of the human intestinal and colon. To assess the genomic stability of our intestinal stem cells, we serially passaged two independent intestinal stem cell pedigrees derived from the ileum of one late fetal demise case and tested them after 50 (passage 5; P5), 100 (P10), 150 (P15) and 200 days (P20) of continuous proliferation. DNA from cells at these passages cells was analyzed by SNP array based copy number variation analysis (CNV). Although at P15 and P20, partial aneuploidy in some of pedigrees was observed, there are almost no CNVs at P5 and P10 in both pedigrees.

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Project description:Stem cells of the gastrointestinal tract, pancreas, liver and other columnar epithelia collectively resist cloning in their elemental states. Here we demonstrate the cloning and propagation of highly clonogenic, 'ground state' stem cells of the human intestine and colon. We show that derived stem-cell pedigrees sustain limited copy number and sequence variation despite extensive serial passaging and display exquisitely precise, cell-autonomous commitment to epithelial differentiation consistent with their origins along the intestinal tract. This developmentally patterned and epigenetically maintained commitment of stem cells is likely to enforce the functional specificity of the adult intestinal tract. Using clonally derived colonic epithelia, we show that toxins A or B of the enteric pathogen Clostridium difficile recapitulate the salient features of pseudomembranous colitis. The stability of the epigenetic commitment programs of these stem cells, coupled with their unlimited replicative expansion and maintained clonogenicity, suggests certain advantages for their use in disease modelling and regenerative medicine.

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