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Adaptation to culture of human embryonic stem cells and oncogenesis in vivo

Abstract

The application of human embryonic stem cells (HESCs) to provide differentiated cells for regenerative medicine will require the continuous maintenance of the undifferentiated stem cells for long periods in culture. However, chromosomal stability during extended passaging cannot be guaranteed, as recent cytogenetic studies of HESCs have shown karyotypic aberrations. The observed karyotypic aberrations probably reflect the progressive adaptation of self-renewing cells to their culture conditions. Genetic change that increases the capacity of cells to proliferate has obvious parallels with malignant transformation, and we propose that the changes observed in HESCs in culture reflect tumorigenic events that occur in vivo, particularly in testicular germ cell tumors. Further supporting a link between culture adaptation and malignancy, we have observed the formation of a chromosomal homogeneous staining region in one HESC line, a genetic feature almost a hallmark of cancer cells. Identifying the genes critical for culture adaptation may thus reveal key players for both stem cell maintenance in vitro and germ cell tumorigenesis in vivo.

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Figure 1: Representative cytogenetic data from a HESC line maintained in Sheffield.
Figure 2: Ideogram of all reported chromosome abnormalities in HESCs.
Figure 3: Interphase FISH analysis of H14.s3 cell line hybridized with an iso17q probe (Kreatech Biotechnology) specific for the p53 (17p13) and MPO (17q23) genes.
Figure 4: Evidence of a minimal region of amplification on chromosome 17q in culture-adapted HESCs.
Figure 5: Characterization of the HSR in the abnormal HESC line H14.

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Acknowledgements

We are grateful to C. Cowan and D. Melton for providing the HUES1-17 HESC lines, and to J. Thomson for the H1, H7 and H14 HESC lines. We are grateful to our colleagues, especially J. Jackson, K. Amps, G. Bray and G. Bingham for culture of the HESCs. In addition, we would like to acknowledge B. Aflatoonian and L. Ruban for their assistance in the derivation and proliferation of the Shef cell lines. This work was supported by grants from the Medical Research Council, Yorkshire Cancer Research, The Engineering and Physical Sciences Research Council and The Juvenile Diabetes Research Foundation.

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Supplementary Figure 1

FISH on the Shef5 cell line with the SMS and MDS probes specific for the critical regions of Smith Magenis syndrome and Miller Dieker syndrome at 17p11.2 and 17p13.3, respectively. (PDF 457 kb)

Supplementary Methods (PDF 132 kb)

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Baker, D., Harrison, N., Maltby, E. et al. Adaptation to culture of human embryonic stem cells and oncogenesis in vivo. Nat Biotechnol 25, 207–215 (2007). https://doi.org/10.1038/nbt1285

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