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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۵۷-۵۷
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عنوان انگلیسی Ps-25: The Impact of Platelet Factor Rich Supernatant as An Effective Alternative Source to Fetal Bovine Serum for Umbilical Cord Blood Mesenchymal Stem Cells Proliferation
چکیده انگلیسی مقاله Objective: Umbilical Cord Blood derived Mesenchymal Stem Cells (UCB-MSCs) are promising candidates for cell-based therapy. Fetal Bovine Serum (FBS) is commonly used as a cell culture medium additive for in vitro UCB-MSCs expansion. However, the use of animal sera for human cell-based culture may be associated with ethical, and safety issues. Among the possible alternatives to the animal serum, platelet derived compounds have been proposed. Materials and Methods: For preparation of PFRS, whole-blood donation of four healthy donors was used to prepare one pooled platelet concentrate out of buffy coats. Then, platelet concentrate was activated by human thrombin and calcium chloride. UCB-MSCs were obtained with informed consent and cultured in Dulbecco’s modified Eagle’s medium-low Glucose (DMEM), supplemented with 10% FBS and characterized with flow-cytometry and differentiated into the adipogenic and osteogenic lineages. Then for comparison, UCB-MSCs passages 4-8 were cultured in expansion medium consisted of DMEM containing either 10% FBS as a control group or 10% PFRS as a test group; then cell viability and population doubling rate for two groups were performed. To assess cell viability, 5000 cells were plated in each well of a 24-well plate and cell viability was then evaluated at different time points (T24h, T48h, T72h, T96h, 120h) via MTT. To determine the cell proliferation and population doubling rate at each passages, 2000 cells per square centimeter were plated in T25 flasks then UCB-MSCs were counted and passaged at a confluence of 80%. Results: MTT results did not show significant increase in cell viability after adding PFRS to the culture medium in compare to FBS. Cell counting in different passages exhibited significant increase in population doubling rate (3.7 ± 0.36 vs. 3 ± 0.4, p≤0.05), and fold expansion (1.6 fold) in culture media containing PFRS. Conclusion: Our results demonstrated the PFRS application as a safe, rich and ethically approved source that can be substituted with FBS in cellular therapy. In addition, functional capability of PFRS would be approved in animal model as well.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/351
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