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대한생식의학회지 제35권 제4호 2010년
사람의 제대 및 양막유래 줄기세포의 간세포로의 분화
서울여자대학교 자연과학대학 생명공학과1, 수원제일산부인과2, 연세대학교 의과대학 외과학교실3, 연세대학교 의과대학 산부인과학교실4
김지영1•이윤정2•박세아1•강현미1•김경식3•조동제4•김해권1*,
Hepatogenic Potential of Umbilical Cord Derived-Stem Cells and Human Amnion Derived-Stem Cells
Jiyoung Kim1, Yoon Jung Lee2, Seah Park1, Hyun Mi Kang1, Kyung Sik Kim3,Dong Jae Cho4, Haekwon Kim1*
1Department of Biotechnology, College of Natural Sciences, Seoul Women's University, Seoul, Korea,2Suwonjeil Women's Hospital, Suwon, Korea, 3Department of Surgery, College of Medicine, Yonsei UniversitySeoul, Korea, 4Department of OB/GYN, College of Medicine, Yonsei University, Seoul, Korea
Objectives: Many types of liver diseases can damage regenerative potential of mature hepatocytes, hepatic progenitor cells or oval cells. In such cases, a stem cell-based therapy can be an alternative therapeutic option. We examined whether human amnion-derived mesenchymal stem cells (HAM) and human umbilical cord-derived stem cells (HUC) could differentiate into hepatocyte-like cells as therapeutic cells for the liver diseases. Methods: HAM and HUC were isolated from the amnion and umbilical cord of the volunteers after a caesarean section with informed consent. In order to differentiate these cells into hepatocyte-like cells, cells were cultivated in hepatogenic medium using culture plates coated with fibronectin. Effects of hepatocyte growth factor, L-ascorbic acid 2-phosphate, insulin premixture fibroblast growth gactor 4, dimethylsulfoxide, oncostatin M and/or dexamethasone were examined on the hepatic differentiation. After differentiation, the cells were analyzed by RT-PCR, immunocytochemistry, immunoblotting, albumin ELISA, urea assay and periodic acid-schiffs staining. Results: Initial fibroblast-like appearance of HAM and HUC changed to a round shape during culture in the hepatogenic medium. However, in all hepatogenic conditions examined, HUC secreted more amounts of albumin or urea into medium than HAM. Expression of some of hepatocyte-specific genes increased and expression of new genes were observed in HUC following cultivation in hepatogenic medium. Results of immunocytochemistry and immunoblotting analyses demonstrated that HUC secreted albumin into the culture medium. PAS staining further demonstrated that HUC could store glycogen inside of the cells. Conclusions: Both HUC and HAM could differentiate into albumin-secreting, hepatocyte-like cells. Under the same hepatogenic conditions examined, HUC more efficiently differentiated into hepatocyte-like cells compared with the HAM. The results suggest that HUC and HAM could be used as sources of stem cells for the cell-based therapeutics such as in liver diseases.
키워드 : Umbilical cord, Amnion, Stem cells, Hepatocyte, Differentiation
교신저자 : hwkim@swu.ac.kr
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