摘要
目的探讨应用脂质体将Hif-1α稳定转染至MSCs是否影响其与β-TCP支架的相容性。方法应用直接贴壁法提取原代MSCs,流式细胞仪鉴定干细胞表面抗原并行向成骨细胞诱导分化鉴定。通过脂质体将Hif-1α转染至第3代(P3)MSCs,经遗传霉素(G418)筛选获得稳定转染的细胞。将β-TCP支架分为两组:A组,MSCs与β-TCP支架共培养;B组,Hif-1α稳定转染的MSCs与β-TCP支架共培养。观察比较两组样品中MSCs的成活细胞数和生长曲线,分析Hif-1a稳定转染对MSCs与β-TCP支架相容性的影响。结果 Hif-1α转染前后MSCs均可在β-TCP支架上良好生长,且两组细胞均保持细胞干性,两组间细胞数目和细胞增殖差异无统计学意义(P>0.05)。结论 Hif-1α转染不会对MSCs在β-TCP支架上的生长增殖产生影响。
Objective To investigate the effect of hypoxia inducible factor-1α （HIF-1α） on compatibility of bone marrow mesenchymal stem cells （MSCs） with beta tricalcium phosphate （β-TCP） scaffold. Methods The primary MSCs were isolated by direct adherent method. Flow cytometry was performed for validation of MSCs as well as its differential capacity to osteoblast. HIF-1α was transfected into the third generation （P3） of MSCs by liposome. HIF-1α positive cells were screened by geneticin 418 （G418）. β-TCP scaffold plus MSCs was divided into 2 groups： group A in which MSCs and β-TCP scaffolds were co-cultured, group B in which HIF-1α positive MSCs and β-TCP were co-cultured. The survival rate and growth curve of MSCs were observed. Result MSCs grew well on the β-TCP scaffolds prior and post HIF-1α transfection, maintaining differential capacity without statistical signifcance （P 〉 0.05）. No signifcant difference was observed between the two groups in cell number and cell proliferation （P 〉 0.05）. Conclusion HIF-1α expression does not affect the growth and proliferation of MSCs on β-TCP scaffold.
出处
《中国现代医学杂志》
CAS
2018年第3期16-20,共5页
China Journal of Modern Medicine
基金
沈阳医学院科技基金(No:20155047)
辽宁省教育厅科技一般项目(No:L2013316)
