Stem Cells and Cloning: Advances and Applications (Nov 2020)
The Potentiality of Human Umbilical Cord Isolated Mesenchymal Stem/Stromal Cells for Cardiomyocyte Generation
Abstract
Amoura Abou-ElNaga,1 Farha El-Chennawi,2 Samar Ibrahim kamel,1 Ghada Mutawa3 1Zoology Department, Faculty of Sciences, Mansoura University, Mansoura 35516, Egypt; 2Clinical Pathology Department, Faculty of Medicine, Mansoura University, Mansoura 35516, Egypt; 3Department of Basic Science, Faculty of Dentistry, Horus University in Egypt (HUE), New Damietta 34518, EgyptCorrespondence: Ghada MutawaDepartment of Basic Science, Faculty of Dentistry, Horus University in Egypt (HUE), International Coastal Road New, Damietta City, Kafr Saad, Damietta Governorate 34518, EgyptTel +201021888628Fax +2-057-328611Email [email protected]: The new therapeutic strategy of managing cardiac diseases is based on cell therapy; it highly suggests the use of multipotent mesenchymal stem/stromal cells (MSCs). MSCs widely used in researches are known to be isolated from bone marrow. However, this research seeks to use a human umbilical cord (HUC) as an alternative source of MSCs. Since HUC Wharton’s jelly (WJ)-isolated MSCs originate as fetal tissue they are highly preferable for their potential advantages over other adult tissues.Methods: The researchers used enzymatic digestion to establish a primary HUC-WJ-isolated MSC line. Then, flow cytometry was used to characterize MSCs and hematopoietic stem cells (HSCs) markers’ expression. In addition, the cardiac differentiation capacity of HUC-WJ-isolated MSCs in vitro was investigated by two protocols. Protocol-1 necessitates the dependence on merely 5-azacytidine (5-Aza), whereas in protocol-2, 5-Aza was supported by basic fibroblast growth factor (BFGF). The comparative study between the two protocols was applied by inspecting the ultrastructure of differentiated cells, measuring RT-PCR mRNA cardiac markers and the quantitative detection of cardiac proteins.Results: HUC-WJ isolated MSCs were expressed by CD90+ve, CD105+ve, CD106+ve, CD45−ve, and CD146−ve. Remarkable TNNT1, NKX2.5, and Desmin mRNA expression and higher quantitative LDH and cTnI were detected by applying protocol-2. This same protocol-2 induced cardiac morphological features that were revealed by identifying cardiomyocyte-like cells and typical sarcomeres.Conclusion: HUC-WJ is proved to be an ethical and effective source of MSCs induced cardiac differentiation, whereas BFGF supports 5-Aza in MSCs-cardiomyocytes differentiation.Keywords: mesenchymal stem/stromal cells, human umbilical cord, Wharton’s jelly, cardiomyocytes, 5-azacytidine, fibroblast growth factor
