| Summary: | 碩士 === 元智大學 === 化學工程與材料科學學系 === 101 === Ex vivo culture system are used to expand and differentiate hematopoietic stem cells (HSCs) to solve the cell shortage in therapeutic application. Fetal bovine serum (FBS) is known to promote proliferation of cell. However, FBS always has the issue of xenogenesis virus contamination, such as mad cow disease. Active platelet rich plasma (PRP) is a novel biotechnological production. PRP contains various growth factors that have beneficial effects on cell growth and tissue regeneration. In this study, we tried to develop the umbilical cord blood-PRP (h-PRP) and porcine-PRP (p-PRP) as a FBS substitute in HSC expansion. First, we isolated and purified HSC and PRP from umbilical cord blood. Second, we activated PRP to release various growth factors by stimulation with thrombin, collagen and calcium chloride. Finally, we compared the effects on HSC expansion and differentiation by basal medium containing with FBS and PRP (5%, 10%, 20%, 30%). Our results showed that after 3-day culture, the increases of the CD34+ cells and CD34+CD38- cells in the different of concentration h-PRP medium were 2.43-, 2.38-, 0.81-, 0.93- and 12.95-, 10.11-, 3.02-, 1.22-fold compare to 10% FBS medium, respectively. In addition, the percentages of the differentiated T-lymphocytes (CD3), B-lymphocytes (CD19), granulocyte (CD33), platelets (CD41a), natural killer cell (CD56) and erythrocytes (CD235a) in the total cultured cells were 5.9%, 1.6%, 91%, 6.55%, 0.9% and 29.6%, respectively. The increase number of the CD41a+/CD61+ cells induced by the medium contain 10% autologous hPRP is 9.32 fold compared to the medium contain 10% FBS. In the meanwhile, the increases of the CD34+ cells in the three kinds of the 10% p-PRP medium were 2.76-, 2.42- and 2.92-fold compare to 10% FBS medium, respectively and the increases of CD34+CD38- cells were 4.83-, 5.04-, 6.69-fold, respectively. Finally, our results demonstrated that both active h-PRP and p-PRP had the potentials to replace FBS in HSC culture. h-PRP and p-PRP can increase the commercial value of umbilical cord blood bank and porcine industry in Taiwan.
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