Differentiation of cardiomyocytes from amniotic fluid‑derived mesenchymal stem cells by combined induction with transforming growth factor β1 and 5‑azacytidine

Jiang,Shan; Zhang,Song

doi:10.3892/mmr.2017.7373

Differentiation of cardiomyocytes from amniotic fluid‑derived mesenchymal stem cells by combined induction with transforming growth factor β1 and 5‑azacytidine

Authors:
- Shan Jiang
- Song Zhang
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Affiliations: Cardiovascular Department, Xin Hua Hospital, Affiliated Hospital of Shanghai Jiaotong University, Shanghai 200092, P.R. China
Published online on: August 28, 2017 https://doi.org/10.3892/mmr.2017.7373
Pages: 5887-5893
Copyright: © Jiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

As a novel type of seed cell, amniotic fluid‑derived mesenchymal stem cells (AFMSCs) are promising for the regeneration of myocardial cells. A focus of cardiovascular regenerative medicine is to improve the efficiency of AFMSC differentiation. The present study replaced the traditional method of AFMSC differentiation with a combined induction method, in order to improve the efficiency of directional differentiation. AFMSCs were obtained from rabbit amniotic fluid samples, and western blot analysis was performed to analyze the expression of octamer‑binding transcription factor 4 (OCT4), and tumorigenicity experiments were conducted. AFMSCs were divided into the following 4 groups: Induction with transforming growth factor β1 (TGFβ1); induction with 5‑azacytidine (5Aza); induction with TGFβ1 and 5Aza combined; and untreated controls. Reverse transcription‑quantitative polymerase chain reaction was performed to analyze the expression of cardiac‑specific GATA binding protein 4 (GATA4), and immunofluorescence was employed to analyze the expression of cardiac troponin T (cTnT). In addition, western blotting was performed to analyze the expression of connexin 43, and transmission electron microscopy was used to observe the ultrastructure of the differentiated cells. AFMSCs exhibited positive OCT4 expression and were not observed to induce tumor development in nude mice. The expression levels of GATA4, cTnT, and connexin 43 in the combined induction group were markedly higher when compared with the remaining groups. Transmission electron microscopy analysis revealed that differentiated cells exhibited myocardial cell characteristics. In conclusion, AFMSCs are multipotent, non‑tumorigenic cells that are capable of differentiating into cardiomyocyte‑like cells. This combined induction method may improve the efficiency of directed differentiation.

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