세포성장억제제를 처리하지 않은 지지세포로부터 인간배아줄기세포 배양법 = Human Embryonic Stem Cell Culture Technique on Mitomycin C-free Feeder Cells using Porous Membrane

다국어 초록 (Multilingual Abstract)

For clinical applications of human embryonic stem cells (hESC), it is critical to develop hESCs culture techniques perfectly excluding animal feeder contamination generated by conventional hESCs culture system. Previously, we have developed novel hESCs culture technique using porous membrane (PM) that not only reduced feeder contamination significantly but also provided isolation of hESCs efficiently. However, feeder cells used for hESCs culture were treated with mitomycin C (MMC) that is not acceptable for clinical application. Therefore, we investigated whether PM system can be further applied for MMC non-treated STO feeder cells. MMC non-treated STO cells were seeded on the bottom of PM and then hESCs were placed on the top of PM. Transfer of hESCs were carried out every 6 days. After 10 passages, pluripotency of hESC was verified through immunostaining of Oct4, SSEA3/4, TRA1-60 and TRA-1-81. According to RT-PCR analysis, the hESC strongly expressed stemness markers such as Nanog and Oct4. Compared to conventional hESC culture technique requiring MMC treatment, PM technique would be more useful to maintain clinical grade hESC by eliminating MMC treatment procedure.

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