摘要
目的观察hBcl-2转染的骨髓基质细胞(MSCs)对体外培养PC12细胞的保护作用,为下一步hBcl-2修饰的MSCs脑内移植治疗奠定基础。方法采用密度梯度离心结合贴壁法分离、培养大鼠MSCs,对培养第3代的MSCs进行鉴定;hBcl-2用脂质体转染MSCs建立MSCs-hBcl-2基因工程细胞;用H2O2建立PC12细胞氧化应激损伤模型;MTT法检测MSCs-hBcl-2基因工程细胞上清对体外培养的PC12细胞活性的影响。结果成功培养出MSCs细胞,免疫组化结果显示培养的细胞CD44、CD71表达阳性,而CD45表达阴性;建立了有稳定高效hBcl-2表达的MSCs-hBcl-2基因工程细胞,并观察到MSCs-hBcl-2基因工程细胞较MSCs更能减轻PC12细胞的氧化损伤(P<0.05)。结论hBcl-2基因修饰的MSCs对PC12细胞的生长、生存有着重要的保护作用。
Objective To construct hBcl-2 Genetical Modified Marrow Stromal Cells and observe their influence on cultured PC12 cells,which laid an important foundation in the treatment of brain injury.Methods Density gradient centrifugalization combined with adherence methods were used to segregate and cultivate rat MSCs.The 3rd generation of MSCs were identified.hBcl-2 was transfected in MSCs.PC12 cells model of oxidative stress injury was established with H2O2.MTT method was used to evaluate PC12 cells survival after cultured with MSCs-hBcl-2 conditional medium.Results MSCs were successfully cultivated.Immunocytochemistry showed that CD44 and CD71 expression was positive while CD45 expression were negative for MSCs.The genetically engineered cells MSCs-hBcl-2 was established and improved the survival rate of the PC12 cells more remarkably than MSCs.Conclusion Human Bcl-2（hBcl-2）gene-modified MSCs play an important role in the protection of the PC12 cell growth and survival.
出处
《脑与神经疾病杂志》
2010年第2期140-142,共3页
Journal of Brain and Nervous Diseases
