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Korean Journal of Fertility and Sterility 2004;31(4):209-216.
Published online December 1, 2004.
Induced Differentiation of Embryonic Stem Cells to Insulin Secreting Cells.
Ji Hye Sung, Chun Kyu Lim, Hye Won Choi, Hyoung Song Lee, Hyeonsang Shin, Jin Hyun Jun, Hyun Soo Yoon, Mi Kyoung Koong
1Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital, Sungkyunkwan University School of Medicine, Seoul, Korea.
2Department of Ob/Gyn Samsung Cheil Hospital, Sungkyunkwan University School of Medicine, Seoul, Korea.
3MizMedi Hospital, Medical Research Center, Seoul, Korea.
Abstract
OBJECTIVE
Embryonic stem (ES) cells could be differentiated into the specific cell types by alternation of culture condition and modification of gene expression. This study was performed to evaluate the differentiation protocol for mouse and human ES cells to insulin secreting cells. METHODS: Undifferentiated mouse (JH-1) and human (Miz-hES1) ES cells were cultured on STO feeder layer, and embryoid bodies (EBs) were formed by suspension culture. For the differentiation, EBs were cultured by sequential system with three stage protocol. The differentiating ES cells were collected and marker gene expressions were analyzed by semi-quantitative RT-PCR in each stage. Amount of secreted insulin levels in culture media of human ES cells were measured by human insulin specific RIA kit. RESULTS: During the differentiation process of human ES cells, GATA-4, alpha-fetoprotein, glucose transporter-2 and Ngn-3 expression were increased whereas Oct-4 was decreased progressively. Insulin and albumin mRNAs were expressed from stage II in mouse ES cells and from stage III in human ES cells. We detected 3.0~7.9 microU/ml secretion of insulin from differentiated human ES cells by in vitro culture for 36 days. CONCLUSION: The sequential culture system could induce the differentiation of mouse and human ES cells into insulin secreting cells. This is the first report of differentiation of human ES cells into insulin secreting cells by in vitro culture with serum and insulin free medium.
Key Words: Embryonic stem cell; Insulin secreting cell; Differentiation; Sequential system; Serum and insulin free medium


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