We achieved possibility of isolation, characterization human umbilical cord blood endothelial progenitor cells (EPCs), examination potency of EPCs to form new blood vessels and differentiation into cardiomyoctes in canines with acute myocardial infarction (AMI). EPCs were separated and cultured from umbilical cord blood. Their phenotypes were confirmed by uptake of double stains dioctadecyl tetramethylindocarbocyanine-labeled acetylated LDL and FITC-labeled Ulex europaeus agglutinin 1 (DILDL-UEA-1). EPCs of cord blood were counted. Human VEGFR-2 and eNOS from the cultured EPCs were assessed by qPCR. Human EPCs was transplanted intramyocardially in canines with AMI. ECG and cardiac enzymes (CK-MB and Troponin I) were measured to assess severity of cellular damage. Histopathology was done to assess neovascularisation. Immunostaining was done to detect EPCs transdifferentiation into cardiomyocytes in peri-infarct cardiac tissue. qPCR for human genes (hVEGFR-2, and eNOS) was done to assess homing and angiogenic function of transplanted EPCs. Cultured human cord blood exhibited an increased number of EPCs and significant high expression of hVEGFR-2 and eNOS genes in the culture cells. Histopathology showed increased neovascularization and immunostaining showed presence of EPCs newly differentiated into cardiomyocyte-like cells. Our findings suggested that hEPCs can mediate angiogenesis and differentiate into cardiomyoctes in canines with AMI.
Chronic renal failure (CRF) is a common disease with high morbidity and mortality. The inadequacy of current treatment modalities and insufficiency of donor organs for cadaveric transplantation have driven a search for improved methods of dealing with renal failure. This raises the concept of cell-based therapy using mesenchymal stem cells (MSCs). The purpose of this work is to evaluate the effect of MSCs in rats with CRF and to detect the level of B-cell lymphoma 2 (Bcl-2) and basic fibroblast growth factor (bFGF) as possible antiapoptotic and anti-inflammatory paracrine mediators of MSCs action. This study was carried on female albino rats, which were divided into 3 groups, control group, CRF group and CRF/MSCs group. MSCs were derived from bone marrow of male albino rats and were characterized morphologically and by RT-PCR for CD29. Serum urea, creatinine, Na, K, systolic blood pressure and 24-hour urinary albumin were estimated for all groups, Y-chromosome gene (Sry) was detected by PCR in renal tissue. Also Bcl-2 and bFGF were examined by RT-PCR in renal tissue of all studied groups and the kidney was examined pathologically. The results of this work showed that CRF rats receiving MSCs showed significantly lowered serum urea, creatinine and urinary albumin levels compared to the CRF group. Also improvement of serum Na and K was observed in CRF/MSCs group compared to CRF group. As regard Bcl-2 and bFGF they were highly expressed in renal tissue of CRF/MSCs group compared to the CRF group. The (Sry) gene was detected by PCR in the renal tissue of CRF/MSCs group. These results demonstrate a potential of MSCs to ameliorate the kidney function in rats with CRF possibly through the antiapoptotic and anti-inflammatory paracrine mediators of MSCs
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