Lack of effect of plasma of myocardial preconditioned, ischemic and ischemic-reperfused rats of myocardium on differentiation of mesenchymal stem cells into cardiomyocytes

Mansour Esmailidehaj; Hassan Esmaeili; Elham Chavoushi; Mohammad Ebrahim Rezvani; Hossein Azizian

doi:10.18502/ssu.v30i4.9902

Mansour Esmailidehaj Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Hassan Esmaeili Pediatric Department, School of Medicine, Golestan University of Medical Sciences, Gorgan, Iran.
Elham Chavoushi Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Mohammad Ebrahim Rezvani Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Hossein Azizian Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.

DOI: https://doi.org/10.18502/ssu.v30i4.9902

Keywords: Mesenchymal stem cells, ischemia preconditioning, plasma, Desmin, β-MHC and troponin I.

Abstract

Introduction: Numerous studies have shown that enriched plasma protects myocardial cells against ischemia. The aim of this study was to evaluate the effect of rat enriched plasma by preconditioning, ischemia, and reperfusion on the differentiation of bone marrow mesenchymal stem cells (BMMSC) into cardiomyocytes.

Methods: In this experimental and laboratory study, BMMSCs were extracted from the femur bone of male Wistar rats weighting 250-300 g. BMMSCs of the third passage were divided into six groups according to the type of plasma received: 1) control, 2) azacytidine, 3) azacytidine and non-ischemic plasma recipient, 4) azacytidine and preconditioned plasma recipient , 5) azacytidine and ischemic plasma recipient 6) azacytidine and plasma reperfusion recipient groups. In all groups except group 1, to initiate cardiomyocyte differentiation, BMMSCs were exposed to azacytidine (10 μM) for 24 hours. Plasma (25 μg / ml) was added to the culture medium each time the culture medium was changed. After 21 days, the expression level of myocardial βMHC, desmin, and troponin I genes was assessed using qRT-PCR.

Results: The expression of myocardial βMHC, desmin, and troponin I genes was significantly increased in all groups received azacytidine (groups 2 to 6) (P<0.01). There was no significant difference in the expression of myocardial βMHC, desmin, and troponin I genes, the doubling time, and migration between the plasma receiving groups (groups 3 to 6) compared to the group 2 (P>0.05).

Conclusion: The findings of present study showed that the plasma collected during ischemia, ischemia- reperfusion, and ischemia preconditioning had no effect on the differentiation of BMMSCs into cardiomyocytes in the presence of azacytidine.