Effect of ascorbic acid on differentiation of human amniotic fluid mesenchymal stem cells into cardiomyocyte-like cells

Runchana Markmee; Sirinda Aungsuchawan; Peraphan Pothacharoen; Waleephan Tancharoen; Suteera Narakornsak; Tanongsak Laowanitwattana; Kanokkan Bumroongkit; Chaniporn Puaninta; Nathaporn Pangjaidee

doi:10.1016/j.heliyon.2019.e02018

Effect of ascorbic acid on differentiation of human amniotic fluid mesenchymal stem cells into cardiomyocyte-like cells

Heliyon. 2019 Jul 17;5(7):e02018. doi: 10.1016/j.heliyon.2019.e02018. eCollection 2019 Jul.

Authors

Runchana Markmee¹, Sirinda Aungsuchawan¹, Peraphan Pothacharoen², Waleephan Tancharoen¹, Suteera Narakornsak¹, Tanongsak Laowanitwattana¹, Kanokkan Bumroongkit¹, Chaniporn Puaninta¹, Nathaporn Pangjaidee¹

Affiliations

¹ Department of Anatomy, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
² Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.

Abstract

The aim of this study was to evaluate the efficiency of ascorbic acid (AA) on cell viability, cytotoxicity and the effects on cardiomyogenic differentiation of the human amniotic fluid mesenchymal stem cells (hAF-MSCs). The results of methylthiazole tetrazolium (MTT) assay and cell apoptosis assay indicated that after 24, 48 and 72 h of treatment, AA had no effect on cells viability and cytotoxicity. After treating the hAF-MSCs with 5-azacytidine (5-aza) and a combination of AA and 5-aza, the alamar blue cells proliferation assay showed the normal growth characteristic similar to control group. Especially, the morphological changes were observed between day 0 and day 21, and it was revealed that the hAF-MSCs exhibited myotube-like morphology after 7 days of cell culturing. Moreover, the treatment with a combination of AA and 5-aza was able to up-regulate the cardiomyogenic specific gene levels, which are known to play an important role in cardiomyogenesis. This was specifically notable with the results of immunofluorescence and immunoenzymatic staining in the AA combined with 5-aza treatment group, the highest expression of cardiomyogenic specific proteins was revealed including for GATA4, cTnT, Cx43 and Nkx2.5. It could be concluded that AA may be a good alternative cardiomyogenic inducing factor for hAF-MSCs and may open new insights into future biomedical applications for a clinically treatment.

Keywords: 5-Azacytidine; Ascorbic acid; Biological sciences; Cardiomyocyte-like cells; Cardiomyogenic differentiation; Cell biology; Human amniotic fluid mesenchymal stem cells; Stem cell research.