Combination of retinoic acid, dimethyl sulfoxide and 5-azacytidine promotes cardiac differentiation of human fetal liver-derived mesenchymal stem cells

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• 作者：Fuxue Deng ; Han Lei ; Yunfeng Hu ; Linjing He ; Hang Fu ; Rui Feng…
• 关键词：Human fetal ; liver derived mesenchymal stem cells ; 5 ; Azacytidine ; Retinoic acid ; Dimethyl sulfoxide ; Cardiomyocyte ; like cells ; Pluripotency ; Oct4
• 刊名：Cell and Tissue Banking
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：17
• 期：1
• 页码：147-159
• 全文大小：7,754 KB
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• 作者单位：Fuxue Deng (1)
  Han Lei (1)
  Yunfeng Hu (1)
  Linjing He (1)
  Hang Fu (1)
  Rui Feng (1)
  Panpan Feng (1)
  Wei Huang (1)
  Xi Wang (1)
  Jing Chang (1)
  
  1. Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Cell Biology
  Biomedicine
  Transplant Surgery
  
• 出版者：Springer Netherlands
• ISSN：1573-6814

文摘

There are controversial reports about cardiac differentiation potential of mesenchymal stem cells (MSCs), and there is still no well-defined protocol for the induction of cardiac differentiation. The effects of retinoic acid (RA) and dimethyl sulfoxide (DMSO) on the proliferation and differentiation of human fetal liver-derived MSCs (HFMSCs) as well as the pluripotent state induced by 5-azacytidine (5-aza) in vitro were investigated. MSCs were isolated from fetal livers and cultured in accordance with previous reports. Cells were plated and were treated for 24 h by the combination of 5-aza, RA and DMSO in different doses. Different culture conditions were tested in our study, including temperature, oxygen content and medium. Three weeks later, cells were harvested for the certification of cardiac differentiation as well as the pluripotency, which indicated by cardiac markers and Oct4. It was found that the cardiac differentiation was only induced when HFMSCs were treated in the following conditions: in high-dose combination (5-aza 50 μM + RA 10−1 μM + DMSO 1 %) in cardiac differentiation medium at 37 °C and 20 % O₂. The results of immunohistochemistry and quantitative RT-PCR showed that about 40 % of the cells positively expressed Nkx2.5, desmin and cardiac troponin I, as well as Oct4. No beating cells were observed during the period. The combined treatment with RA, DMSO and 5-aza in high-dose could promote HFMSCs to differentiate into cardiomyocyte-like cells and possibly through the change of their pluripotent state.