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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۷۸-۷۸
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عنوان انگلیسی Ps-46: Umbilical Cord Blood-Derived CD133+ Stem Cells Generate Small Colony Forming Unit-Megakaryocyte In Vitro: A Hypothetical Reason for Tardiness in Platelet Recovery after Cord Blood Transplantation
چکیده انگلیسی مقاله Objective: Although umbilical cord blood was considered as a waste material, now many cord blood banks save and prepare it for transplantation. The main concern following cord blood transplantation is tardiness of platelet recovery. One of the probable Feasons of this problem is incapacity to generate large colonies of megakaryocytic lineage. This hypothesis may be reasonable since larger colonies can produce more platelet.Materials and Methods: In this study, we evaluated the colonogenic ability of CD133+ derived from cord blood in collagen-based medium in vitro. First, CD133+ cells were purified by direct and indirect magnetic sorting methods. Then the sorted cells were compared in terms of CD133, CD41, CD61, and CD42b expression by flow cytometric analysis immediately after purification and after 7 day culture in Stem Span serum-free-medium. Also, CD133+ and CD133- cells were seeded in collagen-based MegaCult-C medium containing recombinant human insulin, human transferrin, L-glutamine, 2-mercaptoethanol, rh TPO, rh IL- 6, and rh IL-3 at 37°C and humidified condition for at most two weeks. According to manufacturer’s instructions, colonies were stained by biotin-avidin conjugate for CD41 antigen and finally categorized in three groups: small, medium, and large. Results: Our data showed that there was not any significant difference between direct and indirect method in expression of CD41, CD61, and CD42b in both after sorting and at day 7 of culture in Stem Span media. But it appeared that indirect method were more efficient to purify of CD133+ cells. In our experiments, CD133- cells could not generate any megakaryocytic colonies. CD133+ cells formed three categorized colonies though most of them were small. Conclusion: Consequently, indirect magnetic method was more effective for purification of CD133+ cells. Since CD133+ derived from cord blood generated about 64% small colonies, it may convince delayed platelet recovery after cord blood transplantation.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/372
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