自体富血小板血浆凝胶析出液对骨髓间充质干细胞迁移影响的体外研究

The effectc of autologous platelet-rich plasma gel precipitation liquid on the migration of marrow mesenchymal stem cell migration

目的:观察富血小板血浆凝胶(PRG)析出液对骨髓间充质干细胞(BMSCs)迁移的影响。方法:全骨髓贴壁法体外培养犬BMSCs,二次离心法分离提取富血小板血浆并制备其凝胶析出液;将第3代BMSCs接种于Transwell小室,分别用含PRG析出液体积分数为0%(对照组)、1%、5%、10%、20%、50%的DMEM进行培养,观察5%PRG析出液作用不同时间(12、24、36 h)对BMSCs迁移活性的影响。结果:当培养液中含PRG析出液的浓度为1%、5%、10%、20%时,能明显促进BMSCs的迁移(P﹤0.05),而当PRG析出液浓度为50%时,则抑制BMSCs的迁移(P﹤0.05);5%PRG析出液促迁移作用最强,此后随PRG析出液浓度升高,促迁移作用越来越低,各浓度组间两两相比差异均有统计学意义(P<0.05);5%PRG析出液组作用于BMSCs 12、24、36 h均能明显促进BMSCs迁移(P<0.05),12 h组低于24 h和36 h组(P﹤0.05),24 h组与36 h组相比P﹥0.05。结论:含1%、5%、10%、20%PRG析出液的培养基均可明显促进BMSCs的迁移,在一定浓度范围内呈时间依赖性。

AIM： To investigate the effect of Platelet-rich plasma gel （PRG） precipitation liquid （PL） on the migration of bone marrow stem cells （BMSCs） in vitro. METHODS ： Canine BMSCs was obtained by adherence separation method. PRGPL was prepared by duplicatese centrifugation and diluted with DMEM to a volume fraction of 1%, 5%, 10%, 20% and 50% solution, BMSCs were cultured with PRGPL at the various concentrations for 24 h at 0（control） and 5% for 12 h, 24 h and 36 h respectively. The migration of BMSCs was examined by migration assay with transwell migration model. RESULTS： PRGPL at 1% ,5%, 10% and 20% promoted the migration of BMSCs （ P 〈 0.05 ） ; while at the concentration of 50% restrained the migration（ P 〈 0.05 ）. PRGPL at 5% showed the maxi- mum effect and time-dependeant migration increase （ between 12 h and 24 h culture, P 〈 0.05 ）, between 24 h and 36 h culture, P 〉 0.05）. CONCLUSION： The platelet-rich plasma gel precipitation liquid at concentrations of 1%, 5% ,10% ,20% can promote the migration of BMSCs, this migration effect is in a time-dependent manner.