title: Combination of retinoic acid, dimethyl sulfoxide and 5-azacytidine promotes cardiac differentiation of human fetal liver-derived mesenchymal stem cells.
abstract: 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.
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Fuxue
authors.initials: F
authors.lastname: Deng
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Han
authors.initials: H
authors.lastname: Lei
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Linjing
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Hang
authors.initials: H
authors.lastname: Fu
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Feng
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Panpan
authors.initials: P
authors.lastname: Feng
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China. jingchangdoc@126.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Chang
mesh_terms: D001374:Azacitidine / Q000494:pharmacology*; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D048430:Cell Shape / Q000187:drug effects; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005333:Fetus / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000196:embryology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009206:Myocardium / Q000166:cytology; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D010641:Phenotype; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D015415:Biomarkers; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D014212:Tretinoin; D001374:Azacitidine; D004121:Dimethyl Sulfoxide
keywords: 5-Azacytidine; Cardiomyocyte-like cells; Dimethyl sulfoxide; Human fetal-liver derived mesenchymal stem cells; Oct4; Pluripotency; Retinoic acid