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BOR - Papers in Press, published online ahead of print August 18, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.033480
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Submitted June 24, 2004
Returned for revision July 19, 2004
Accepted August 17, 2004

Reproductive Technology


Establishment and Maintenance of Human Embryonic Stem Cell Lines on Human Feeder Cells Derived from Uterine Endometrium under Serum-Free Condition

Jung Bok Lee , Jeoung Eun Lee , Jong Hyuk Park , Sun Jong Kim , Moon Kyoo Kim , Sung Il Roh , and Hyun Soo Yoon *

* To whom correspondence should be addressed. E-mail: yoon{at}mizmedi.net.

Abstract
Human embryonic stem (hES) cells are usually established and maintained on mouse embryonic fibroblasts (MEFs) feeder layers. However, it is desirable to develop human feeder cells because animal feeder cells are associated with risks such as viral infection and/or pathogen transmission. In this study, we attempted to establish new hES cell lines using human uterine endometrial cells (hUECs) to prevent the risks associated with animal feeder cells and for their eventual application in cell replacement therapy. Inner cell masses (ICMs) of cultured blastocysts were isolated by immunosurgery and then cultured on mitotically inactivated hUEC feeder layers. Cultured ICMs formed colonies by continuous proliferation and were allowed to proliferate continuously for 40, 50, and 55 passages. The established hES cell lines (Miz-hES14, -15, and -9, respectively) exhibited typical hES cells characteristics including continuous growth, expression of specific markers, normal karyotypes, and differentiation capacity. hUEC feeders have the advantage that they can be used for many passages, whereas MEF feeder cells can only be used as feeder cells for a limited number of passages. hUECs are available to establish and maintain hES cells, and the high expression of embryotrophic factors and extracellular matrices by hUECs may be important to the efficient growth of hES cells. Clinical applications require the establishment and expansion of hES cells under stable xeno-free culture systems.

Key words: Embryo


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