Cardiomyocyte-like cells differentiation from non β-catenin expression mesenchymal stem cells
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Recent studies have shown that block wnt/β-catenin signaling pathway is integrant for cardiomyocytes differentiation from bone marrow mesenchymal stem cells (MSCs). By transducing the MSCs with lentivirus which contain β-catenin interference RNA, we screened out the non β-catenin expression clone. In the establishment of knockdown β-catenin in MSCs, we investigated the role of 5-azacytidine (5-aza), salvianolic acid B (salB), and cardiomyocytes lysis medium (CLM) in inducing MSCs to differentiate into cardiomyocyte-like cells. A method for culturing MSCs and cardiomyocytes was established. Purified MSCs were investigated by flow cytometry. The MSCs were positive for CD90 and CD29, but negative for CD34 and CD45. Meanwhile, the cardiomyocytes contracted spontaneously after 24 h of seeding into the plates. The fourth-passage non-β-catenin expression MSCs were divided into eight groups: control group, 5-aza, salB, CLM, 5-aza + salB, 5-aza + CLM, salB + CLM, and 5-aza + salB + CLM. The gene and protein expression of cTnT, α-actin, β-myosin, β-catenin, and GSK-3β were detected by quantitative real-time PCR and Western blotting. Our results showed that cTnT expression in 5-aza + salB + CLM group was ninefold higher than in the control group in the non-β-catenin MSCs model, implying that cardiomyocytes differentiation from MSCs is an extremely complicated process and it is necessary to consider the internal and external environmental conditions, such as suitable pharmaceutical inducers, cardiomyocytes microenvironments, inhibition of the negative signaling pathway and so on.
KeywordsMesenchymal stem cells Cardiomyocytes Differentiation Wnt/β-catenin signaling
This project was supported by the National Natural Science Foundation of China (No. 81173413, www.nsfc.gov.cn), and the Specialized Research Fund for the Doctoral Program of Higher Education of China (Nos. 20101210110004 and 20101210120008, www.cutech.edu.cn), and Tianjin Research program of Application Foundation and Advanced Technology (No. 09JCYBJC12200, www.tsyc.gov.cn). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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