不同浓度同种异体富血小板血浆提取液对脂肪干细胞体外生长状况的影响

发布时间：2018-01-07 19:08

本文关键词：不同浓度同种异体富血小板血浆提取液对脂肪干细胞体外生长状况的影响　出处：《南昌大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：观察不同浓度的同种异体富血小板血浆（Platelet-rich plasma, PRP）提取液对脂肪干细胞（Adipose-derived stem cells,ADSCs）体外生长状况的影响，为将同种异体PRP提取液应用于创面修复提供实验依据。方法：取SD大鼠腹股沟脂肪组织进行原代、传代培养；通过将获得的细胞向脂肪、骨和神经方向诱导分化，鉴定其干细胞特性；三次离心法制备PRP提取液，并加入DMEM/F12配制体积分数分别为6.67%、3.35%、1.67%的培养液；将第3代ADSCs分为4组：A、B、C组分别以含体积分数为6.67%、3.35%、1.67%PRP提取液的培养液干预ADSCs生长，，D组(对照组)ADSCs用含10%胎牛血清的完全培养基培养； A、B、C、D四组细胞分别培养24h、48h后观察ADSCs生长状态，并用MTT法测定细胞增殖活性（OD值）。用SPSS17.0统计软件分析数据。结果：原代培养获得贴壁生长细胞，呈成纤维细胞样，并成功向脂肪细胞、骨细胞和神经细胞方向诱导分化，即具有多向分化潜能，证明所培养的细胞为ADSCs； PRP中血小板浓度为309.26×1010/L，全血中的血小板浓度为621.05×10~9/L，PRP中血小板浓度约为全血的5倍；MTT法测定OD值显示：A、B、C、D组在培养24h后OD值分别为0.4343±0.0843、0.3516±0.0766、0.2581±0.0600、0.2592±0.0736，在培养48h后OD值分别为1.0168±0.4436、0.7417±0.1097、0.3678±0.0342、0.3957±0.1062，A组和B组OD值高于C组或D组，A组OD值高于B组，组间比较差异有统计学意义(P0.05)，但C组OD值和D组相比较差异无统计学意义(P0.05)。结论：通过胶原酶消化、原代培养的方法可获得大量ADSCs；适当浓度的同种异体PRP提取液可显著促进ADSCs体外增殖，有望将同种异体PRP提取液应用于创面修复。
[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 卤0.0843 3 卤0.3516 卤0.0766 after 24 hours of culture. The OD values of 0.2581 卤0.0600,0.2592 卤0.0736 were 1.0168 卤0.4436 卤0.7417 卤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 卤0.0342 卤0.3957 卤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.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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