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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۴۴-۴۴
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عنوان انگلیسی Ps-56: The Effects of Bone Morphogenetic Protein-4 on Cardiomyocyte Differentiation of Mouse
چکیده انگلیسی مقاله Objective: The aim of the present study was to evaluate the effects of BMP-4 on cardiomyocyte differentiation from mouse adipose tissue-derived stem cells (ADSCs). Materials and Methods: ADSCs were isolated from inguinal adipose tissue of 8 to 10-week old mice. Third-passaged ADSCs were cultured to reach 100% confluency. Cardiac differentiation was initiated in DMEM plus 10% fetal bovine serum (FBS) or 15% Knockout Serum Replacement (KoSR). BMP-4 with the final concentrations of 10, 20 or 50 ng/ml was added to each experimental setting for 5 days. After this initial stage, differentiation was continued in FBS-containing medium, and cardiac differentiation was examined after 21 days. Optimization of the differentiation by insulin-transferrin-selenium (ITS) and 5-azacytidine was also investigated. The expression of cardiac transcription factors and cardiac-specific genes was evaluated by RT-PCR. MLC2V and MLC2A mRNAs were compared between groups by quantitative real- time PCR. Moreover, the presence of cardiac-specific proteins was assessed by immunocytochemistry. Results: Three-week differentiated ADSCs showed the expression of cardiac transcription factors, GATA4 and MEF2C, and cardiac-specific genes, α-MHC, β-MHC, MLC2V, MLC2A and ANP. MLC2V mRNA had its highest expression when the initial phase of differentiation was performed in KoSR-containing DMEM and at the presence of 50 ng/ml BMP-4. The highest expression of MLC2A mRNA was detected at the presence of KoSR and 10 ng/ml BMP-4. Supplementing the culture medium of 10 ng/ml BMP-4 group with ITS upregulated the expression of MLC2A mRNA, while neither ITS nor ITS plus 5-azacytidine could increase the expression of MLC2V. Immunocytochemical analyses showed the expression of α-actinin and cardiac troponin I proteins in the ADSC-derived cardiomyocytes. Conclusion: The results of the present study are indicating an important role for BMP-4 in cardiomyocyte differentiation from ADSCs. Moreover, cardiac differentiation was more effective when the initial phase of differentiation was performed in KoSR-containing media.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/860
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