title: Efficient culture system for human embryonic stem cells using autologous human embryonic stem cell-derived feeder cells.
abstract: Human embryonic stem cells (hESCs) need feeder cells for their maintenance in an undifferentiated state. In conventional culture systems, mouse embryonic fibroblasts (MEFs) serve as feeder cells to maintain hESCs. However, the use of MEFs elevates the risk of transmitting mouse pathogens and thus limits the potential of hESCs in cell replacement therapy. Consequently, the use of human feeder cells would be an important step forward in this in vitro technology. To address this issue, we used fibroblast-like cells differentiated from the Miz-hES6 hESC line (DiffMiz-hES6) as feeder cells to support the in vitro growth of three hESC lines. Immunofluorescence microscopy and reverse transcription-PCR assessing the expression of undifferentiated hESC markers revealed all three hESC lines were maintained in an undifferentiated state. In vitro proliferation proceeded as efficiently as when the hESCs were cultured on MEFS. Moreover, karyotype analysis revealed the chromosomal normality of the hESC lines and the DiffMiz-hES6 feeders themselves after even 50 passages. Furthermore, the hESC lines maintained their pluripotency since they remained capable of forming embryoid bodies (EBs) in vitro. Thus, hESC-derived fibroblast-like cells successfully support in vitro hESC propagation.
authors.affiliation: Laboratory of Stem Cell Research, Medical Research Center, MizMedi Hospital, 701-4 Naebalsan-dong, Kangseo-gu, Seoul 157-280, Korea.
authors.forename: Seung Jun
authors.initials: SJ
authors.lastname: Yoo
authors.forename: Byung Sun
authors.initials: BS
authors.lastname: Yoon
authors.forename: Jin Mee
authors.initials: JM
authors.lastname: Kim
authors.forename: Ji Min
authors.initials: JM
authors.lastname: Song
authors.forename: Sung ll
authors.initials: Sl
authors.lastname: Roh
authors.forename: Seungkwon
authors.initials: S
authors.lastname: You
authors.forename: Hyun Soo
authors.initials: HS
authors.lastname: Yoon
mesh_terms: D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007621:Karyotyping; D039904:Pluripotent Stem Cells / Q000166:cytology; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors
chemical_list: D015415:Biomarkers