Clinical and Experimental Reproductive Medicine

  • 제34권4호
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  • Pages.239-252
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  • 2007
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  • 2233-8233(pISSN)
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  • 2233-8241(eISSN)
대한생식의학회 (The Korean Society for Reproductive Medicine)
권호 표지

사람의 지방 및 제대에서 유래된 유사중간엽 줄기세포로부터 심근세포로의 분화 유도

Cardiomyogenic Potential of Human Adipose Tissue and Umbilical Cord Derived-Mesenchymal Like Stem Cells

  • 박세아 (서울여자대학교 자연과학대학 생명공학과) ;
  • 강현미 (서울여자대학교 자연과학대학 생명공학과) ;
  • 김은수 (서울여자대학교 자연과학대학 생명공학과) ;
  • 김진영 (서울여자대학교 자연과학대학 생명공학과) ;
  • 김해권 (서울여자대학교 자연과학대학 생명공학과)
  • Park, Se-Ah (Department of Biotechnology, College of Natural Sciences, Seoul Women's University) ;
  • Kang, Hyeon-Mi (Department of Biotechnology, College of Natural Sciences, Seoul Women's University) ;
  • Kim, Eun-Su (Department of Biotechnology, College of Natural Sciences, Seoul Women's University) ;
  • Kim, Jin-Young (Department of Biotechnology, College of Natural Sciences, Seoul Women's University) ;
  • Kim, Hae-Kwon (Department of Biotechnology, College of Natural Sciences, Seoul Women's University)
  • 발행 : 2007.12.31
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초록

목 적: 사람의 HAD와 HUC를 심근세포로 분화 유도하고자 하였다. 연구방법: 사람의 HAD와 HUC를 분리하여 5-azacytidine을 24시간 처리하고 여러 가지 BMP와 FGF을 첨가하여 배양하였다. 또한 HUC은 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1 또는 Wnt inhibitor를 첨가하여 배양한 후 심근세포 특이 유전자의 발현을 조사하였다. 결 과: HAD를 5-azacytidine 처리하고 기본배양액에서 4주 동안 배양하였을 때 TnT 유전자가 새로이 발현하였으며 Cmlc1과 kv4.3의 발현 양이 증가하였다. 5-azacytidine 처리 후에 BMP-4와 함께 FGF-4 (B4/F4) 또는 FGF-8 (B4/F8)을 첨가하여 배양하였을 때는 $\beta$-MHC 유전자 발현이 새로이 유도되었으며, Cmlc1, TnT, TnI 그리고 Kv4.3 유전자 발현 양이 더 많이 증가하였다. HUC은 5-azacytidine 및 BMP와 FGF 처리에 의해 유전자 발현 변화가 없었다. 그러나 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1을 첨가하여 배양하였을 때, BMP-2와 FGF-8 (B2/F8)을 첨가하여 배양한 세포에서 $\beta$-MHC 발현이 새로이 유도되었으며 $\alpha$-CA, TnT 그리고 Kv4.3 유전자의 발현이 증가하였다. 또한 BMP와 FGF와 함께 Wnt inhibitor를 처리하여 1주 동안 배양하였을 때 Cinlc1 유전자 발현이 새로이 유도되었으며 $\alpah$-CA, TnT, TnI 그리고 Kv4.3의 발현이 증가되었다. 결 론: HAD는 BMP와 FGF 처리에 의해 심근세포 특이 유전자의 발현증가를 유도할 수 있었으며 HUC는 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1 또는 Wnt inhibitor를 처리함으로써 심근세포 특이 유전자의 발현증가를 유도할 수 있었다. 따라서 HAD와 HUC는 심장질환 치료를 목적으로 하는 세포 치료에 이용될 수 있을 것으로 사료된다.

Objectives: In the present study, we examined the differentiation potential of human adipose-(HAD) and human umbilical cord-derived mesenchymal like stem cells (HUC) into cardiomyocytes. Methods: Cells were initially exposed to 5-azacytidine for 24h cells and then were cultivated in the presence or absence of activin A, TGF-$\beta$1, or Wnt inhibitor with various combinations of BMP and FGF. Assessment of cardiomyogenic differentiation was made upon the expression of cardiomyocyte-specific genes using RT-PCR. Results: HAD that cultivated in control medium for 4 weeks after 5-azacytidine expose showed new expression of TnT gene and increased expression of Cmlc1 and kv4.3 genes. However, HAD cultivated in the presence of combinations of BMP-4/FGF-4 (B4/F4) and BMP-4/FGF-8 (B4/F8) showed new expression of $\beta$-MHC gene and more increased expression of Cmlc1, TnT, TnI, Kv4.3 genes. Significantly enhanced expression of Cmlc1, TnT, and Kv4.3 genes were also observed compared to that cultivated in the control medium. Treatment of HUC with either 5-azacytidine or combinations of BMP and FGF did not affect the expression profile of these genes. However, when activin A or TGF-$\beta$1 was present in addition to the BMP-2/FGF-8 (B2/F8) after 5-azacytidine exposure, HUC exhibited new expression of $\beta$-MHC gene and increased expression of $\alpha$-CA, TnT and Kv4.3 genes. When Wnt inhibitor was present in addition to BMP and FGF, HUC showed new expression of Cmlc1 gene and increased expression of $\alpha$-CA, TnT, TnI and Kv4.3 genes. Conclusions: Based on these observations, it is suggested that HAD and HUC could differentiate into cardiomyocytes which might be used as therapeutic cells for the heart diseases.

키워드

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