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대한생식의학회지 제31권 제4호 2010년
배아줄기세포의 인슐린 분비세포로의 유도 분화에 대한 연구
성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실1,산부인과2,미즈메디병원 의과학연구소3
성지혜1, 임천규1,최혜원1,이형송1,신현상1,전진현1*,윤현수3,궁미경2
Induced Differentiation of Embryonic Stem Cells to Insulin Secreting Cells
Ji Hye Sung1,Chun Kyu Lim1,Hye Won Choi1,Hyoung Song Lee1, Hyeonsang Shin1,Jin Hyun Jun1*, Hyun Soo Yoon3, Mi Kyoung Koong2
1Laboratory of Reproductive Biology & Infertility,2Department of Ob/Gyn Samsung Cheil Hospital, Sungkyunkwan University School of Medicine, Seoul, Korea 3MizMedi Hospital, Medical Research Center, Seoul, Korea
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, α-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 μU/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.
키워드 : Embryonic stem cell, Insulin secreting cell, Differentiation, Sequential system, Serum and insulin free medium *본 연구는 과학기술부 21세기 프론티어 연구개발 사업단인 세포응용연구사업단의 연구비 지원 (SC12022)으로 수행되었습니다.
교신저자 : junjh55@hanmail.net
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