الفهرس 
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Abstract
Mesenchymal stem cell (MSC) transplantation has been proposed as a promising therapeutic strategy for ischemic myocardium repair following myocardial infarction. Differentiation of MSCs into cardiomyocyte-like cells prior to cell transplantation is advantageous in improving their potential clinical benefits for cardiac repair.
MSCs are the spindle shaped plastic-adherent cells isolated from bone marrow, adipose, and other tissue sources, with multipotent differentiation capacity in vitro. These cells can differentiate into a variety of cell types, Including: osteoblasts (bone cells) , chondrocytes (cartilage cells) , Myocytes (muscle cells) ,And adipocytes (fat cells). This phenomenon has been documented in specific cells and tissues in living animals and their counterparts growing in tissue culture.
Bone marrow Mesenchymal stem cells have been proposed as potential sources of stem cells for regeneration of the CVS , With the advantage of autologous transplantation which avoids the immune rejection and ethical concerns, MSCs have great application prospect in personalized treatment of cardiovascular diseases.
The aim of this work is to study cardiogenesis using AZA for induction of BMSCs differentiation and then follow the cells as they form cardiomyocytes. The present study was carried out on 30 patients referred to clinical pathology department, El- Menoufia University Hospital between May 2015 to May 2017 .
MNCs separated from the samples using ficoll-hypaque (1:2 ratio), suspended in DMEM-media, The MSCs suspensions were seeded at concentration of (million cells/ ml) in tissue culture plastic flasks 25 cm2 incubated in 5 ml of fresh nutrient medium for 9 days, At day 9 adherent cells were harvested by trypsinization and identified by flowcytometric analysis of CD73,CD44 and CD34 expression.
The MSCs differentiation into cardiac cell induced by treatment with 5-azacytidine , Basic fibroblast growth factor (bFGF) and Insulin-like Growth Factor I (IGF-I) .It was proven by morphology and flowcytometric expression of cardiac connexin and vimentin.
The results of this study revealed successful isolation of MSCs from bone marrow aspirate ,MSCs identified by morphology, CD44 expression CD73 expression as positive marker and lack of specific cell surface markers of hematopoietic cells like CD34. MSCs showed positive expression for CD 44 (ranging between 67.10-97.50 with a mean ± standard deviation of 83.01 ±9.55)and showed positive expression for CD 73 (ranging between 53.40 – 95.30 with a mean ± standard deviation of 75.38 ± 11.44)While they showed negative expression for CD 34 (ranging between 0.10-2.0, with a mean ± standard deviation of 0.92 ±0.55).
Induced MSCs with 5-AZA were regularly monitored using phase-contrast microscope and inverted microscope, these cells showed morphological changes consistent with cardiogenesis as compared to the symmetric morphology of the unindicted cells.
Induced cells showed high positive expression of vimentin with a range of(62.80 – 95.30) and mean ± standard deviation of 82.46 ±9.05 and a range of (62.70 – 93.20) and mean ± standard deviation of 80.0 ± 9.06 for connexin.