本文關鍵詞：不同濃度同種異體富血小板血漿提取液對脂肪干細胞體外生長狀況的影響　出處：《南昌大學》2012年碩士論文　論文類型：學位論文

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【摘要】：目的： 觀察不同濃度的同種異體富血小板血漿（Platelet-rich plasma, PRP）提取液對脂肪干細胞（Adipose-derived stem cells,ADSCs）體外生長狀況的影響，為將同種異體PRP提取液應用于創(chuàng)面修復提供實驗依據(jù)。 方法： 取SD大鼠腹股溝脂肪組織進行原代、傳代培養(yǎng)；通過將獲得的細胞向脂肪、骨和神經(jīng)方向誘導分化，鑒定其干細胞特性；三次離心法制備PRP提取液，并加入DMEM/F12配制體積分數(shù)分別為6.67%、3.35%、1.67%的培養(yǎng)液；將第3代ADSCs分為4組：A、B、C組分別以含體積分數(shù)為6.67%、3.35%、1.67%PRP提取液的培養(yǎng)液干預ADSCs生長，，D組(對照組)ADSCs用含10%胎牛血清的完全培養(yǎng)基培養(yǎng)； A、B、C、D四組細胞分別培養(yǎng)24h、48h后觀察ADSCs生長狀態(tài)，并用MTT法測定細胞增殖活性（OD值）。用SPSS17.0統(tǒng)計軟件分析數(shù)據(jù)。 結(jié)果： 原代培養(yǎng)獲得貼壁生長細胞，呈成纖維細胞樣，并成功向脂肪細胞、骨細胞和神經(jīng)細胞方向誘導分化，即具有多向分化潛能，證明所培養(yǎng)的細胞為ADSCs； PRP中血小板濃度為309.26×1010/L，全血中的血小板濃度為621.05×10~9/L，PRP中血小板濃度約為全血的5倍；MTT法測定OD值顯示：A、B、C、D組在培養(yǎng)24h后OD值分別為0.4343±0.0843、0.3516±0.0766、0.2581±0.0600、0.2592±0.0736，在培養(yǎng)48h后OD值分別為1.0168±0.4436、0.7417±0.1097、0.3678±0.0342、0.3957±0.1062，A組和B組OD值高于C組或D組，A組OD值高于B組，組間比較差異有統(tǒng)計學意義(P0.05)，但C組OD值和D組相比較差異無統(tǒng)計學意義(P0.05)。 結(jié)論： 通過膠原酶消化、原代培養(yǎng)的方法可獲得大量ADSCs；適當濃度的同種異體PRP提取液可顯著促進ADSCs體外增殖，有望將同種異體PRP提取液應用于創(chuàng)面修復。
[Abstract]:Objective: Platelet-rich plasma was observed in platelet-rich plasma with different concentrations of allogeneic platelet-rich plasma. Effects of PRP extract on the growth of Adipose-derived stem cells in vitro. To provide experimental basis for the application of allogenic PRP extract in wound repair. Methods: The inguinal adipose tissue of SD rats was subcultured. The cells were induced to differentiate into fat, bone and nerve to identify their stem cell characteristics. The PRP extract was prepared by three times centrifugation, and the volume fraction of DMEM/F12 was 6.67% and 3.35% respectively. The third generation of ADSCs was divided into 4 groups: group C: ADSCs growth was interfered with 6.67% and 3.35% and 1.67% respectively. Group D (control group) was cultured on a complete medium containing 10% fetal bovine serum. The growth state of ADSCs was observed after cultured for 24 h or 48 h. The OD value of proliferative activity was measured by MTT and the data were analyzed by SPSS17.0 software. Results: Adherent growth cells were obtained in primary culture, which were fibroblast-like, and were successfully induced to adipocytes, bone cells and nerve cells, that is, multidirectional differentiation potential. The cultured cells were proved to be ADSCs. The platelet concentration in PRP was 309.26 脳 1010 / L, and the platelet concentration in whole blood was 621.05 脳 10 ~ (-9) / L ~ (-1). The OD value measured by MTT method showed that the OD value of group D was 0.4343 3 鹵0.0843 3 鹵0.3516 鹵0.0766 after 24 hours of culture. The OD values of 0.2581 鹵0.0600,0.2592 鹵0.0736 were 1.0168 鹵0.4436 鹵0.7417 鹵0.1097 after 48 h culture, respectively. The OD values of group A and group B were higher than those of group C or group D (0.3678 鹵0.0342 鹵0.3957 鹵0.1062). There was significant difference between group C and group D, but there was no significant difference in OD value between group C and group D (P 0.05). Conclusion: Through collagenase digestion, the primary culture method can obtain a large number of ADSCs; The proper concentration of allogenic PRP extract can significantly promote the proliferation of ADSCs in vitro, which is expected to apply allogenic PRP extract to wound repair.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R329

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