Transforming growth factor-&beta;1 combined with tanshinoneⅡA induces bone marrow mesenchymal stem cells differentiating into cardiomyocyte-like cells in vitro

Lü Yang LIU Bo WANG Hai-ping* LIU Yuan SUN Wei LI Rou CHEN Xiao-yi

doi:10.16098/j.issn.0529-1356.2016.05.007

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Acta Anatomica Sinica ›› 2016, Vol. 47 ›› Issue (5) : 620-627. DOI: 10.16098/j.issn.0529-1356.2016.05.007

Transforming growth factor-β1 combined with tanshinoneⅡA induces bone marrow mesenchymal stem cells differentiating into cardiomyocyte-like cells in vitro

LÜ Yang¹ LIU Bo² WANG Hai-ping ^1* LIU Yuan¹ SUN Wei¹ LI Rou¹ CHEN Xiao-yi¹

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Abstract

Objective The present study aims to test the hypotheses that if transforming growth factor beta 1(TGF-β1) or tanshinoneⅡA or their combination enhance the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) towards the cardiomyogenic phenotype, and TGF-β1 combined with tanshinoneⅡA may achieve better effects than the alone group. Methods BMSCs were isolated from bones of limbs of 10 male SD rat and cultured. The 2nd-generation BMSCs were co-incubated with TGF-β1 or tanshinoneⅡA or their combination for 72 hours. The control group was BMSCs which cultured without any inductive substance. The morphological characteristics, surface antigens and mRNA expression of several transcription factors in each group were assessed. Results BMSCs were initially spindle-shaped with irregular processes. The cells were gradually increased after been inoculating for 24 hours and proliferated 1 week later. On week 4, BMSCs in the combined group showed fusiform shape, orientating with one accord, and were connected with adjoining cells forming myotube-like structures. The morphous of BMSCs in the alone induced group was similar to that in the combined group, but the amount of the cells was relatively low. Immunohistochemistry revealed the expressions of the cardiac-specific markers, tropomyosin, Cx43 and troponin Ⅰ, in the combined group were significantly higher than that of the other groups (P<0.05). The transcriptional expressions of the cardiomyocyte-specific genes GATA-4,Nkx2.5 were highest in 1week after induction, and they were all stable and significantly higher in the induced groups than that in the control group. Transmission electron microscope showed that the induced cells had a cardiomyocyte-like ultrastructure: the nucleus was round and located in the center of the cells, mitochondria and rough endoplasmic reticula were founded in the cytoplasm, and the paralleled myofilaments were seen in the cytoplasm. Conclusion Taken together, these results indicate that TGF-β1 or tanshinoneⅡA may induce BMSCs to acquire cardiogenic phenotype and TGF-β1 combined with tanshinoneⅡA may achieve better effects than the alone group.

Key words

Bone marrow mesenchymal stem cell / Transforming growth factor beta 1 / TanshinoneⅡA / Cardiomyocyte / Flow cytometry / Immunohistochemistry / Real-time PCR / Rat

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Lü Yang LIU Bo WANG Hai-ping LIU Yuan SUN Wei LI Rou CHEN Xiao-yi. Transforming growth factor-β1 combined with tanshinoneⅡA induces bone marrow mesenchymal stem cells differentiating into cardiomyocyte-like cells in vitro[J]. Acta Anatomica Sinica. 2016, 47(5): 620-627 https://doi.org/10.16098/j.issn.0529-1356.2016.05.007

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