低氧诱导因子-1α 转染对骨髓间充质干细胞与β- 磷酸三钙支架相容性的影响研究

Effect of HIF-1α expression on compatibility of MSCs with β-TCP scaffolds

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。

Abstract:

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 significance (P >0.05). No significant 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.