Establishment and maintenance of human embryonic stem cell lines on human feeder cells derived from uterine endometrium under serum-free condition.
Human embryonic stem (hES) cells are usually established and maintained on mouse embryonic fibroblast (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-hES-14, -15, and -9, respectively) exhibited typical hES cells characteristics, including continuous growth, expression of specific markers, normal karyotypes, and differentiation capacity. The 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. The 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.
Lee
Jung Bok
JB
Division of Stem Cell Biology, Medical Research Center, MizMedi Hospital, 157-280 Seoul, Korea.
Lee
Jeoung Eun
JE
Park
Jong Hyuk
JH
Kim
Sun Jong
SJ
Kim
Moon Kyoo
MK
Roh
Sung Il
SI
Yoon
Hyun Soo
HS
D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D016895:Culture Media, Serum-Free; D016172:DNA Fingerprinting; D004622:Embryo, Mammalian / Q000166:cytology*; D004717:Endometrium / Q000166:cytology*; D005260:Female; D006028:Glycosphingolipids / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D008297:Male; D051379:Mice; D016513:Mice, SCID; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology; D055746:Stage-Specific Embryonic Antigens; D013234:Stem Cells / Q000166:cytology*; D013724:Teratoma / Q000473:pathology
D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; D016895:Culture Media, Serum-Free; D006028:Glycosphingolipids; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4