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Separation and maintenance of normal cells from human embryonic stem cells with trisomy 12 mosaicism

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Abstract

Human embryonic stem cells (hESCs) are pluripotent and hold great promise as useful tools in basic scientific research and in the field of regenerative medicine. However, several studies have recently reported chromosomal abnormalities such as gains of chromosomes 12, 17 and X in hESCs. This genetic instability presents an obstacle in the application of hESCs as sources of cell therapies. We found that trisomy 12 was correlated with changes in hESC colony morphology during hESC maintenance. In this study, we investigated whether normal and trisomy 12 cells could be separated in hESC cultures displaying trisomy 12 mosaicism with two types of colony morphology using a mechanical transfer technique. Eight sublines were cultured from eight hESC colonies displaying normal or abnormal morphology. Four sublines with normal morphology had normal chromosome 12 numbers, whereas the four sublines with abnormal morphology displayed trisomy 12. These results indicate that a hESC colony with a minor degree of chromosomal mosaicism and normal morphology could proceed to a colony with normal chromosomes after prolonged cultures with mechanical transfer. Therefore, analysis of cultures for chromosomal abnormalities when changes in colony morphology are observed during culture is essential for maintaining normal hESC lines.

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Abbreviations

AP:

alkaline phosphatase

EC:

embryonic carcinoma

FISH:

fluorescence in-situ hybridization

hESC:

human embryonic stem cell

IVF:

in-vitro fertilization

SCID:

severe combined immunodeficiency

SNUhES4:

Seoul National University human Embryonic Stem cell line 4

SSEA-4:

stage specific embryonic antigen-4

TGCT:

testicular germ cell tumour

Tra-1-60:

tumour rejection antigen-1-60

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Acknowledgements

This research was supported by a grant (SC1150) from the Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Education, Science and Technology, Republic of Korea.

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Authors and Affiliations

  1. Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, 110-810, Korea

    Hye Won Seol, Sun Kyung Oh, Hee Sun Kim, Jin Ah Baek, Jin Seo, Eun Hee Kim, Seung Yup Ku, Seok Hyun Kim, Young Min Choi & Shin Yong Moon

  2. Department of Obstetrics and Gynecology, Seoul National University College of Medicine, 28 Yeongeon-dong, Jongro-gu, Seoul, 110-799, Korea

    Sun Kyung Oh, Hee Sun Kim, Seung Yup Ku, Seok Hyun Kim, Young Min Choi & Shin Yong Moon

  3. Central Research Institute, Sam Jin Pharm. Co. Ltd., Hwasung, 445-746, Korea

    Yong Bin Park

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  1. Hye Won Seol
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  2. Sun Kyung Oh
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  3. Yong Bin Park
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  4. Hee Sun Kim
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  9. Seok Hyun Kim
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  11. Shin Yong Moon
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Correspondence to Young Min Choi.

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Seol, H.W., Oh, S.K., Park, Y.B. et al. Separation and maintenance of normal cells from human embryonic stem cells with trisomy 12 mosaicism. Chromosome Res 16, 1075–1084 (2008). https://doi.org/10.1007/s10577-008-1258-y

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  • DOI: https://doi.org/10.1007/s10577-008-1258-y

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