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J Microsc Ultrastruct. 2019 Oct-Dec;7(4):185-193. doi: 10.4103/JMAU.JMAU_17_19. Epub 2019 Nov 18.

Cardiogenic Differentiation of Murine Bone Marrow-Derived Mesenchymal Stem Cells by 5-Azacytidine: A Follow-up In vitro Study.

Author information

1
Department of Histology and Cell Biology, Faculty of Medicine, Tanta University, Tanta, Egypt.
2
Department of Clinical Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt.

Abstract

Background:

Cell-based therapy is a promising tool in the management of myocardial infarction.

Aim of the Work:

The aim of this study is to examine the in vitro potential differentiation of murine bone marrow (BM)-derived stem cells into cardiomyocytes using 5-azacytidine after 1, 3, and 5 weeks and follow it up after 8 weeks.

Materials and Methods:

BM-derived mesenchymal stem cells (MSCs) were extracted from the bones of adult albino rats. MSCs were induced with 10 μM 5-azacytidine for 24 h. The cells were examined after 1, 3, 5, and 8 weeks. Cell characterization with immunocytochemistry for detection of CD105, desmin, and T-troponin and transmission electron microscopy was performed.

Results:

The 5-azacytidine-induced MSCs showed light and electron microscopic histological characteristics resembling cardiomyocytes and progressively expressed the cardiac muscle-specific markers over the 1st, 3rd, and 5th weeks, yet by the 8th week, these parameters were significantly downregulated.

Conclusion:

Prolonged survival of 5-azacytidine-induced MSCs in culture beyond the 8th week resulted in loss of the newly acquired cardiomyocyte characteristics. It is not recommended to prolong the maintenance of 5-azacytidine-induced MSCs in culture on the hope of increasing its cardiogenic potentiality beyond 5 weeks.

KEYWORDS:

5-Azacytidine; cardiogenic differentiation; in vitro; mesenchymal stem cells

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