Available human feeder cells for the maintenance of human embryonic stem cells.
Mouse embryonic fibroblasts (MEFs) have been previously used as feeder cells to support the growth of human embryonic stem cells (hESCs). In this study, human adult uterine endometrial cells (hUECs), human adult breast parenchymal cells (hBPCs) and embryonic fibroblasts (hEFs) were tested as feeder cells for supporting the growth of hESCs to prevent the possibility of contamination from animal feeder cells. Cultured hUECs, hBPCs and hEFs were mitotically inactivated and then plated. hESCs (Miz-hES1, NIH registered) initially established on mouse feeder layers were transferred onto each human feeder layer and split every 5 days. The morphology, expression of specific markers and differentiation capacity of hESCs adapted on each human feeder layer were examined. On hUEC, hBPC and hEF feeder layers, hESCs proliferated for more than 90, 50 and 80 passages respectively. Human feeder-based hESCs were positive for stage-specific embryonic antigen (SSEA)-3 and -4, and Apase; they also showed similar differentiation capacity to MEF-based hESCs, as assessed by the formation of teratomas and expression of tissue-specific markers. However, hESCs cultured on hUEC and hEF feeders were slightly thinner and flatter than MEF- or hBPC-based hESCs. Our results suggest that, like MEF feeder layers, human feeder layers can support the proliferation of hESCs without differentiation. Human feeder cells have the advantage of supporting more passages than when MEFs are used as feeder cells, because hESCs can be uniformly maintained in the undifferentiated stage until they pass through senescence. hESCs established and/or maintained under stable xeno-free culture conditions will be helpful to cell-based therapy.
Lee
Jung Bok
JB
Division of Stem Cell Biology, Medical Research Center, MizMedi Hospital 701-4, Kangseo-ku, Seoul 157-280, Korea.
Song
Ji Min
JM
Lee
Jeoung Eun
JE
Park
Jong Hyuk
JH
Kim
Sun Jong
SJ
Kang
Soo Man
SM
Kwon
Ji Nie
JN
Kim
Moon Kyoo
MK
Roh
Sung Il
SI
Yoon
Hyun Soo
HS
D000328:Adult; D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers / Q000032:analysis; D001940:Breast / Q000166:cytology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques; D004622:Embryo, Mammalian / Q000166:cytology; D004717:Endometrium / Q000166:cytology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D006028:Glycosphingolipids / Q000032:analysis; D006801:Humans; D007621:Karyotyping; D051379:Mice; D055746:Stage-Specific Embryonic Antigens; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000276:immunology
D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; D006028:Glycosphingolipids; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4