Exp Mol Med.  2005 Oct;37(5):399-407.

Efficient culture system for human embryonic stem cells using autologous human embryonic stem cell-derived feeder cells

• Yoo SJ
• Yoon BS
• Kim JM
• Song JM
• Roh SI
• You S
• Yoon HS

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 (Diff (Miz-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 Diff (Miz-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.

Keyword

cell culture techniques; humans; stem cells; stem cell transplantation

MeSH Terms

Biological Markers/analysis
Cell Culture Techniques/*methods
Cell Differentiation
Cell Proliferation
Cells, Cultured
Embryo/*cytology
Fibroblasts/cytology
Humans
Karyotyping
Pluripotent Stem Cells/cytology
Research Support, Non-U.S. Gov't
Stem Cells/*cytology
Time Factors