自体激活富血小板血浆干预兔骨髓间充质干细胞体外成软骨分化的研究

背景：自体富血小板血浆激活后可释放多种生长因子,可以促进骨髓间充质干细胞的增殖与分化。目的：观察自体激活富血小板血浆对体外培养的兔骨髓间充质干细胞向成软骨细胞分化的影响。方法：取兔股骨骨髓,全骨髓贴壁法分离培养骨髓间充质干细胞;取第3代骨髓间充质干细胞,分别应用体积分数10%自体激活富血小板血浆和体积分数10%胎牛血清培养液进行体外培养,观察其向成软骨细胞分化情况。结果与结论：分离培养的兔骨髓间充质干细胞呈长梭形,传代后细胞生长迅速。流式细胞仪检测发现第3代细胞高表达CD29、CD44,而低表达CD45。免疫荧光细胞化学染色显示经自体激活富血小板血浆诱导的骨髓间充质干细胞表达Ⅱ型胶原;实时荧光定量PCR检测发现经自体激活富血小板血浆诱导的骨髓间充质干细胞Ⅱ型胶原α1链基因和聚集蛋白聚糖基因表达明显高于经胎牛血清诱导的骨髓间充质干细胞（P〈0.01）。可见自体激活富血小板血浆具有促进兔骨髓间充质干细胞向软骨细胞方向分化的潜能。

Effect of activated autologous platelet-rich plasma on chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells in vitro

BACKGROUND： Platelet-rich plasma, when activated, could secret multiple growth factors which may promote the proliferation and differentiation of bone marrow-derived mesenchymal stem cells.OBJECTIVE： To explore the effect of activated autologous platelet-rich plasma on the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells in vitro.METHODS： Rabbit bone marrow-derived mesenchymal stem cells were isolated using the whole bone marrow adherence method. Passage 3 bone marrow-derived mesenchymal stem cells were in vitro cultured with 10% activated autologous platelet-rich plasma and 10% fetal bovine serum separately. Chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells in vitro was observed.RESULTS AND CONCLUSION： Bone marrow-derived mesenchymal stem cells were successfully isolated and exhibited a long-shuttle-shaped appearance. Cells grew faster when passaged. Immunofluorescence staining showed that after induced by activated autologous platelet-rich plasma, bone marrow-derived mesenchymal stem cells exhibited type Ⅱ collagen expression. Quantitative real-time PCR analysis showed that the expression of α1 chain of type Ⅱ collagen and aggrecan mRNA was significantly up regulated in the bone marrow-derived mesenchymal stem cells induced by activated autologous platelet-rich plasma than in the cells induced by fetal bovine serum （P 〈 0.01）. Our findings in this study suggested that activated autologous platelet-rich plasma can promote the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells in vitro.