复合富血小板血浆培养液对兔骨髓间充质干细胞增殖及矿化的影响

Effect of PRP - supplemented culture medium on proliferation and mineralization of bone marrow mesenchymal stem cells

目的研究富血小板血浆(platelet-rich plasma,PRP)对于骨髓间充质干细胞(bone marrow mesenchymal stemcells,BMSC)增殖及矿化能力的影响。方法改良Appel法采集兔PRP,同时培养该兔BMSC,将培养成功的BMSC分为实验组及对照组,实验组采用含10%PRP的培养液,对照组采用常规培养液,分别对两组细胞进行细胞增殖的检测(MTT法)、ATP活性检测(试剂盒)及矿化能力检测(Von-Kossa染色)。收集结果并进行统计。结果实验组兔BMSC胞体丰满,细胞突起明显增长;实验组细胞增殖能力强于对照组(P<0.01);实验组细胞在第4、8、12天时ALP活性强于对照组(P<0.05),第16天时无显著差别;实验组每孔矿化结节数(4.083±2.022)个,对照组每孔矿化结节数(3.417±1.285)个,两组比较无统计学差异。结论 PRP对BMSC的生长增殖有促进作用,但对该细胞的矿化能力无明显促进作用。

Objective To investigate the effect of platelet - rich plasma （PRP） on the proliferation and mineralization of bone marrow mesenchymal stem cells （BM -MSCs）, thereby elucidating the possible application of PRP in bone tissue engineering. Meth- ods Rabbit bone marrow aspirates were procured by modified Appel＇ s method. BM - MSCs were isolated and cultured in traditional medium with or without supplement of 10% PRP. The cell proliferation was assayed by MTI＇, cell osteogenic differentiation was as- sayed by using ATP activity examination kit and von - Kossa staining. Results The BM - MSCs cultured in PRP - supplemented me- dium were more spindle - like in morphology and had a significantly shorter doubling time in comparison with the control traditional cul- ture without PRP supplement （P 〈0.01 ）. The cells also displayed a higher ATP activity than control on days 4, 8, 12, and 16 （P 〈 0.05）. No statistically significant difference was exhibited at day 20 post - induction in ATP activity, and von- Kossa staining. Con- clusions PRP enhances the proliferation of BM - MSCs and improves their ATP activity but has no effect on mineralization.