Establishment and maintenance of human embryonic stem cells on STO, a permanently growing cell line.
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.
Park
Jong Hyuk
JH
Division of Stem Cell Biology, Medical Research Center, MizMedi Hospital, Kangseo-ku, Seoul 157-280, Korea.
Kim
Sun Jong
SJ
Oh
Eun Jeong
EJ
Moon
Shin Yong
SY
Roh
Sung Il
SI
Kim
Chul Geun
CG
Yoon
Hyun Soo
HS
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000954:Antigens, Surface; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D004268:DNA-Binding Proteins / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006023:Glycoproteins / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D051379:Mice; D008810:Mice, Inbred C57BL; D050814:Octamer Transcription Factor-3; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D014157:Transcription Factors
D000954:Antigens, Surface; D015295:Antigens, Tumor-Associated, Carbohydrate; D015415:Biomarkers; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D006028:Glycosphingolipids; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C494174:Pou5f1 protein, mouse; D011509:Proteoglycans; D055746:Stage-Specific Embryonic Antigens; C107064:TRA-1-60 antigen, human; D014157:Transcription Factors; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4; D000469:Alkaline Phosphatase