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BOR - Papers in Press, published online ahead of print August 20, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.017467
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BIOLOGY OF REPRODUCTION 69, 2007–2014 (2003)
DOI: 10.1095/biolreprod.103.017467
© 2003 by the Society for the Study of Reproduction, Inc.

Embryo

Establishment and Maintenance of Human Embryonic Stem Cells on STO, a Permanently Growing Cell Line1

Jong Hyuk Park3,4, Sun Jong Kim3,4, Eun Jeong Oh3, Shin Yong Moon5, Sung Il Roh3, Chul Geun Kim4, and Hyun Soo Yoon2,3

Division of Stem Cell Biology,3 Medical Research Center, MizMedi Hospital, Kangseo-ku, Seoul 157-280, Korea Department of Life Science,4 College of Natural Sciences, Hanyang University, Seongdong-gu, Seoul 133-791, Korea Stem Cell Research Center,5 Seoul, Korea

Human embryonic stem (hES) cells have been traditionally cultured on primary mouse embryonic fibroblasts (PMEFs). However, though STO cells have some advantages over PMEFs and human embryonic fibroblasts (hEFs) as feeder cells, they have never been used as feeder cells to establish hES cell lines. In this study, three hES cell lines (Miz-hES1, Miz-hES2, and Miz-hES3) were established from inner cell masses (ICM), using STO as feeder cells. The three hES cell lines had normal karyotypes and expressed high levels of alkaline phosphatase (AP), cell surface markers (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), and transcription factor Oct-4. After culture on STO cells for 2 yr, hES cells maintained the potential to form derivatives of all three embryonic germ layers. Our results show that STO feeder cells have the potential to support the establishment and maintenance of hES cell lines. In addition, our results suggest that laminin may play an important role in maintaining the undifferentiated proliferation of hES cells.

1 Supported by MizMedi Hospital and a grant (code: M102KL010001-02K1201-00310) from the Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology, Republic of Korea.

2 Correspondence: Hyun Soo Yoon, Division of Stem Cell Biology, Medical Research Center, MizMedi Hospital, 701-4 Naebalsan-dong, Kangseo-ku, Seoul, Korea. FAX: 82 2 2007 1285; yoon{at}mizmedi.net




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