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The effect of sodium valproate on differentiation of human adipose-derived stem cells into cardiomyocyte-like cells in two-dimensional culture and fibrin scaffold conditions

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Abstract

Differentiation of mesenchymal stem cells (MSCs) into cardiomyocytes is a complex phenomenon, and attempts to find an effective inducing agent are still ongoing. We studied the effect of fibrin scaffold and sodium valproate (VPA, as a histone deacetylase inhibitor) on the differentiation of human adipose-derived stem cells (hADSCs) into cardiomyocyte-like cells. The cells were cultured in culture flask (2D) and in fibrin scaffold (3D), fabricated of human plasma fibrinogen, with and without VPA (1 mM). QRT-PCR, Western blot, and immunochemistry assays were used to evaluate the expression of cardiac markers at gene and protein levels. High levels of CD44, CD90, CD73, and CD105 were expressed on the surface of hADSCs. Treated encapsulated hADSCs (3D) presented significantly higher mRNA expression of HAND1 (1.54-fold), HAND2 (1.59-fold), cTnI (1.76-fold), MLC2v (1.4-fold), Cx43 (1.38-fold), βMHC (1.34-fold), GATA4 (1.48-fold), and NKX2.5 (1.66-fold) in comparison to 2D conditions at four weeks after induction. The protein expressions of NKX2.5 (0.78 vs 0.65), cTnI (1.04 vs 0.81), and Cx43 (1.11 vs 1.08) were observed in the differentiated cells both in 3D and 2D groups, while control cells were absolutely negative for these proteins. The frequency of cTnI and Cx43-positive cells was significantly higher in 3D (24.2 ± 15 and 12 ± 3%) than 2D conditions (19.8 ± 3 and 10 ± 2%). Overall, the results showed that VPA can increase cardiomyogenesis in hADSCs and that fibrin scaffold enhances the inductive effect of VPA. Results of this study may improve cell-based protocols for implementation of more successful cardiac repair strategies.

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Acknowledgments

This study was granted by the Physiology Research Center of Kerman University of Medical Sciences (Grant No KMU-1395/476). The authors are grateful for the help of Amirkabir University, Tehran, for electron microscopy of the samples.

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Authors and Affiliations

  1. Cardiovascular Research Center, Institute of Basic and Clinical Physiology Sciences and Department of Physiology, Afzalipour Medical Faculty, Kerman University of Medical Sciences, Kerman, Iran

    Hamid Najafipour

  2. Department of Clinical Biochemistry, Faculty of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

    Zahra Bagheri-Hosseinabadi

  3. Pharmaceutics Research Centre, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

    Touba Eslaminejad

  4. Department of Medical Microbiology, and Physiology Research Center, Afzalipour Medical Faculty, Kerman University of Medical Sciences, Kerman, Iran

    Hamid Reza Mollaei

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  1. Hamid Najafipour
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Correspondence to Zahra Bagheri-Hosseinabadi.

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Najafipour, H., Bagheri-Hosseinabadi, Z., Eslaminejad, T. et al. The effect of sodium valproate on differentiation of human adipose-derived stem cells into cardiomyocyte-like cells in two-dimensional culture and fibrin scaffold conditions. Cell Tissue Res 378, 127–141 (2019). https://doi.org/10.1007/s00441-019-03027-5

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  • DOI: https://doi.org/10.1007/s00441-019-03027-5

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