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兔富血小板血浆对定向诱导骨髓间充质干细胞向成骨细胞分化的影响

Effect of autologous platelet-rich plasma on directional induced differentiation of rabbit bone marrow mesenchymal stem cells

摘要目的 研究兔富血小板血浆(PRP)作为血清替代物诱导骨髓间充质干细胞(BMSCs)向成骨细胞分化,观察细胞形态及分化率.方法 取6只兔动脉血制备PRP,从兔髂骨取骨髓BMSCs,分为PRP组、胎牛血清组、无血清组.分别采用10%PRP、10%胎牛血清、无血清,配比DMEM-F12培养.将每组第2代细胞分为实验组和对照组,实验组加入诱导因子地塞米松、β-甘油磷酸钠、维生素C,对照组仍使用原培养基.观察诱导前后各组细胞形态学及各诱导组成骨细胞分化效率差异.结果 PRP组和胎牛血清组细胞诱导前生长迅速,呈长梭形,诱导后细胞体积逐渐增大,形态变成三角形、多角形、圆形.PRP及胎牛血清组平均诱导7d,细胞自发聚集成复层,形成可由茜素红S染成红色的钙结节及类似骨质结构.无血清组平均诱导14d,仅3只兔出现上述成骨表现.诱导20 d终止实验,PRP和胎牛血清组成骨细胞分化效率比较,差异无统计学意义(P>0.05),但两组均优于无血清诱导组,差异有统计学意义(P<0.05).结论 自体PRP可作为血清替代物,用于BMSCs细胞增殖和诱导成骨细胞分化培养.

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abstractsObjective To study the cell morphology and differentiation efficiency when rabbit bone marrow mesenchymal stem cells (BMSCs) were induced osteogenic differentiation as culturing by autologous platelet-rich plasma (PRP) instead of serum,and to explore a new method of inducing BMSCs osteogenic differentiation.Methods The PRP was prepared by arterial blood of rabbit.Punctured and The bone marrow was sampled from rabbit's iliac bone,and BMSCs were collected,which divided into PRP group,fetal calf serum (FBS) group and serum-free control group,and cultured in 10% autologous PRP,10% FBS and serum-free respectively,combined with DMEM-F12 medium.The second generation cells were divided into experimental and control groups.The experimental groups' medium was added dexamethasone,β-sodium glycerophosphate and ascorbic acid,and the control groups went on.The cell morphological difference of each group was Observed between anterior and after inducing differentiation,and compared between each group.Results BMSCs of PRP and FBS groups grew quickly,presented like fusiform form before induction,and increasd in volume,became a triangle,polygonal and round form gradually after osteogenic induction.Cells of PRP and FBS groups aggregated spontaneously and multilayered,and formed calcium nodules and bone-like structure after induced 7 days averagely,which could be stained red by alizarin red S;cells of serum-free groups were induced 14 days averagely,only three samples showed osteogenesis performance.Cells of PRP and FBS groups differentiation efficiency was superior to serum-free groups when inducd 20 days,the difference was statistically significant (P<0.05),and the difference between efficiency of PRP and FBS groups was not significant (P>0.05).Conclusions Autologous PRP could be used to proliferate and induce osteogenic differentiation of BMSCs instead of serum.

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