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Differences between karyotypically normal and abnormal human embryonic stem cells.

ABSTRACT: OBJECTIVES:To compare different biological characteristics of human embryonic stem cells (HESCs) between those with normal and those with abnormal karyotype. MATERIALS AND METHODS:Culture-adapted HESCs (chHES-3) with abnormal karyotype were compared with karyotypically normal cells, with regard to pluripotency and differentiation capacity, ultrastructure, growth characteristics, gene expression profiles and signalling pathways. RESULTS:We found a new abnormal karyotype of HESCs. We observed that chHES-3 cells with normal and abnormal karyotypes shared similarities in expression markers of pluripotency; however, karyotypically abnormal chHES-3 cells had a tendency for differentiation towards ectoderm lineages and were easily maintained in suboptimal culturing conditions. Abnormal chHES-3 cells displayed relatively mature cell organelles compared to normal cells, and karyotypically abnormal chHES-3 cells had increased survival and population growth. Genes related to cell proliferation and apoptosis were up-regulated, but genes associated with genetic instability (p53, Rb, BRCA1) were down-regulated in the karyotypically abnormal cells. CONCLUSION:Karyotypically abnormal chHES-3 cells had a more developed capacity for proliferation, resistance to apoptosis and less genetic stability compared to normal chHES-3 cells and may be an excellent model for studying and characterizing initial stages that determine transition of embryonic stem cells into cancer stem cells.

SUBMITTER: Yang S

PROVIDER: S-EPMC6495872 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature

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Differences between karyotypically normal and abnormal human embryonic stem cells.

Yang S S Lin G G Tan Y-Q YQ Deng L-Y LY Yuan D D Lu G-X GX

Cell proliferation 20100601 3

<h4>Objectives</h4>To compare different biological characteristics of human embryonic stem cells (HESCs) between those with normal and those with abnormal karyotype.<h4>Materials and methods</h4>Culture-adapted HESCs (chHES-3) with abnormal karyotype were compared with karyotypically normal cells, with regard to pluripotency and differentiation capacity, ultrastructure, growth characteristics, gene expression profiles and signalling pathways.<h4>Results</h4>We found a new abnormal karyotype of H ...[more]

PMID: 20546238

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Project description:We recently reported that overexpression of progastrin (PG) in embryonic epithelial cells (HEKmGAS cells) increased proliferation of the cells compared to that of control HEKC cells. Here, we report the novel finding that tumorigenic and metastatic potential of HEKmGAS cells is also increased significantly compared to that of HEKC cells. Cell surface-associated annexinA2 (CS-ANXA2) binds PG and is overexpressed on cancer cells, allowing us to successfully use fluorescently labeled PG peptide for enumerating metastatic lesions of transformed/cancer cells in vivo. Next, we examined the hypothesis that increased tumorigenic/metastatic potential of isogenic HEKmGAS versus HEKC cells maybe due to transformed phenotype of stem cells. FACSorting/FACScanning of cells demonstrated significant increases in percent doublecortin-CAM-kinase-like1 (DCLK1)/Lgr5-positive stem cells, coexpressing cluster of differentiation44 (CD44)/CS-ANXA2, in HEKmGAS versus HEKC cells. Distinct differences were noted in the morphology of HEKC versus HEKmGAS spheroidal growths on nonadherent cultures (selective for stem cells). HEKC spheroids were rounded with distinct perimeters (e.g., basement membranes), whereas HEKmGAS spheroids were amorphous with no perimeters. Relative levels of DCLK1/Lgr5/CD44 and ANXA2/?-catenin/pNF?Bp65/metalloproteinases were significantly increased in HEKmGAS versus HEKC cells, growing as monolayer cultures, 3D spheroids (in vitro), or xenografts (in vivo). Interestingly, HEKC cells enriched for CS-ANXA2 developed amorphous spheroids, whereas downregulation of ANXA2 in HEKmGAS clones resulted in loss of matrixmetalloproteinases (MMPs) and re-formation of rounded spheroids, suggesting that high levels of CS-ANXA2/MMPs may impact spheroid morphology. Downregulation of DCLK1 significantly attenuated activation of ?-catenin, with loss of proliferation of HEKmGAS and HEKC cells, suggesting that DCLK1 is required for maintaining proliferation of cells. Our results suggest the novel possibility that transformed stem cells, unlike nontransformed stem cells, coexpress stem cell markers DCLK1 and CD44 with CS-ANXA2.

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Differences between karyotypically normal and abnormal human embryonic stem cells.

Publications

Differences between karyotypically normal and abnormal human embryonic stem cells.

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