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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۲۶-۲۶
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عنوان انگلیسی Ps-15: An Efficient Method for Isolation and Identification of Ovine Fetus Bone Marrow Mesenchymal Stem Cells
چکیده انگلیسی مقاله Objective: Mesenchymal stem cells (MSCs) are vastly used in different disease cell therapies, like cardiovascular disease. Most research is done on small model animals particularly mice. However significant differences exist between small animals and human. For this reason, we developed an efficient method of MSC isolation from ovine fetus which does not have the aging problems of adult MSCs. Materials and Methods: Bone marrow aspirations were isolated from different bones like humorous, femur and tibia separately. MSCs were isolated in Ficol gradient from bone marrow aspirations and cultured in DMEM medium. The forth passage-cells were analyzed with CD44 and CD34 markers to identify MSCs by Flow cytometry. Also we used CHEMICON’s Alkaline Phosphatase Detection Kit. Results: Flow cytometric analysis revealed that the cells expressed high levels of human matrix markers. They were positive for CD44 and negative for CD34. We applied alkaline phosphatase to detect stem cells and observed that colonies expressed alkaline phosphatase. In femur and humorous samples, there were cells identified as monocytes by IHC. They omitted MSCs from culture medium, so to separate them from each other, we used gelatin coated plates. Conclusion: To identify ovine MSCs, we can use anti human MSC markers instead of anti ovine MSC markers. Also to get more homogenous MSCs without monocytes, it is better to use tibia bone marrow. We suggest that using gelatinated plates is the optimal condition for separating MSCs and monocytes.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/820
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