摘要
目的建立前强啡肽（PDP）基因修饰的人骨髓间充质干细胞株并验证其可在体外增强细胞强啡肽的分泌。方法本实验运用载体AdSF35-PDP转染人骨髓间充质干细胞hBMSCs，获得分泌强啡肽的干细胞株（hBMSCs-PDP），以转染空载体的细胞（hBMSCs-空载体）和未转染的细胞hBMSCs作为对照，此3组细胞分别采用流式细胞仪检测细胞表面抗原CD29、CD44、CD34、CD45的表达，免疫荧光法检测强啡肽的表达，RT—PCR法检测强啡肽mRNA的表达、WesternBlot法测定强啡肽蛋白的分泌。结果体外实验以流式细胞仪检测此3组细胞的表面标志物CD29、CD44表达均阳性，CD34、CD45表达均阴性且这3组细胞的表达差异比较无统计学意义（P〉0．05）。与hBMSCs和hBMSCs-空载体比较，hBMSCs—PDP细胞株强啡肽mRNA的表达和强啡肽蛋白的表达均上调（P〈0．05）。结论本实验成功构建了前强啡肽基因修饰的人骨髓间充质干细胞，并证明转染后的细胞株hBMSCs—PDP可稳定分泌强啡肽并保留干细胞原有的特征表型。
Objective To establish the human bone marrow mesenchymal stem cell line genetically modified by prodynorphin gene. Methods The hBMSCs cells was transfected with the adenovirus vector Ad5F35-PDP and got the cell line hBMSCs-PDP, the empty vector was control group. The superficial markers of the cells were detected by flow cytometer and PCR, the expression of dynorphin was determined by Western Blot respectively. Results The flow cytometer results showed that all the three different groups hBMSCs cells had positive expression on CD29 and CD44 and negative expression on CD34 and CD45. Compared with hBMSCs-empty vector and hBMSCs, the protein expression of dynorphin was significantly increased in the hBMSCs-PDP cells （P〈 0.05）. Conclusion A human mesenchymal stem cell line that expresses dynorphin is successfully established and the intrinsical characteristics of hBMSCs is invariant.
出处
《实用疼痛学杂志》
2012年第4期247-250,共4页
Pain Clinic Journal
基金
2010年广东省科技科学研究基金（20108031600153）、广东省自然科学基金（10151008004000007）
关键词
强啡肽
腺病毒
干细胞
Dynorphin
Adenovirus
Stem cells
