عزل الخلايا الجذعية المتوسطة من نسيج الحبل السري و تنميطها و تمايزها

The last decades witnessed an increasing interest in stem cells as an alternative source for injured tissues and lost cells either physiologically or morbidly during the adult’s life. Today, few new umbilical cord (UC) blood and tissue banks have been established in Syria aiming at storing these tissues as a source for the future use of hematopoietic stem cells (HSCs) or mesenchymal stem cells (MSCs) found in the cord tissues. This research aims at establishing and testing protocols for isolation, in vitro proliferation, phenotyping, and differentiation of MSCs embedded in umbilical cord tissues of Syrian newborns. MSCs were isolated from two caesarian births depending on their adherence characteristic on plastic surfaces, and cells were cultured in MSC medium for their growth and proliferation. Cell phenotyping was performed by flow cytometry using fluorescent monoclonal antibodies specific to MSCs’ surface markers. Cultured cells were passed several times and a portion of these cells was cryopreserved in liquid nitrogen, and cell viability was assessed. Differentiation of these MSCs into adipocytes was conducted using culture medium Indomethacin and hydrocortisone. In this study we were able to isolate MSCs from umbilical cord blood using standard protocols. These cells grew in culture as adherent cells and exhibited a profile of surface antigens (CD90 ، + CD105+ ،CD13+ ،CD117 ، + CD44 ، + CD34-/dim) specific to MSCs. Indomethacin and hydrocortisone induced differentiation of cultured cells into adipocytes. Cell differentiation was accompanied by changes in cell morphology and emergence of lipid droplets that stained positively with Sudan Oil. Finally, MSCs were successfully cryopreserved in liquid nitrogen with average viability of approximately 90%. To the best of our knowledge, this work is the first of its kind in Syria in which MSCs were isolated from umbilical cord tissues. We are hopeful that this study will pave the way for future research centered on stem cells that constitutes one of the pillars for regenerative medicine and treatment of numerous diseases