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Copy number variation of human intestinal stem cells with different passage in culture (five pedifrees, passage 5 and 25)

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE66748 | GEO | 2015/05/29

SECONDARY ACCESSION(S): PRJNA277856

REPOSITORIES: GEO

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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.

2015-05-29 | GSE66747 | 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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Custom array CGH validation of de novo CNVs in CEU HapMap samples

Project description:This is the validation data for candidate de novo CNV calls made in the CEU Hapmap by Itsara et al., Genome Research 2010. In this study, de novo CNV calls were initially made with Illumina 1M SNP arrays. Validation of CNV calls was performed with Nimblegen custom array CGH using the extended CEPH pedigrees. A truly de novo CNV would be unobserved in the first generation (CEU trio parents), validated in the second generation (CEU trio children), and assuming no selective effects, transmitted to approximately half of the individuals in the third generation. We attempted validation of 4 de novo CNVs in 3 extended CEPH pedigrees: 1358, 1408, and 1459. 12 samples were hybridized in each of the three pedigrees (36 samples total) against a previously well-characterized reference (GM15510; see Tuzun et al., Nat Genet 2005).

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Custom array CGH validation of de novo CNVs in CEU HapMap samples

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Homo sapiens

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Copy number variation analysis of human Barrett's esophagus stem cells (HumanOmniZhongHua-8 v1.1 Beadchip)

Project description:Barrett’s esophagus confers significant risk of esophageal adenocarcinoma. We have established the cloning system of patient-matched stem cells of Barrett’s esophagus and gastric cardia. Barrett's esophagus (BE) stem cells and gastric cardia (GC) stem cells from 12 patients were cloned. To analyze copy number variation in BE and GC stem cells, we have performed SNP array. It has shown that deletions such as p16 and FHIT in BE stem cells are significantly detected, while amplifications in BE stem cells are not. Also, we found some of BE stem cells did not share these deletions, suggesting emerging of BE does not require specific CNV.

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