新型化学转染试剂对人脐带间充质干细胞的转染优化

Optimizing transfection of human umbilical cord-derived mesenchymal stem cells using novel chemical transfection reagents

背景:人脐带间充质干细胞的基因修饰技术多种多样,它们在效率、可靠性及安全性方面各有不同。目的:对比3种新型化学转染试剂在人脐带间充质干细胞中的瞬时转染效率。方法:采用FugeneHD、LipofectamineLTX及Attractene分别转染人脐带间充质干细胞,荧光显微镜下观察阳性细胞,流式细胞术分析不同转染试剂的转染效率,锥虫蓝染色观察转染后人脐带间充质干细胞的生存率。结果与结论:LipofectamineLTX转染效率最高,显著高于FugeneHD和Attractene[(32.50±2.12)%,(4.30±0.64)%,(1.70±0.08)%,P<0.05]。LipofectamineLTX转染后的细胞生存率略低于FugeneHD及Attractene,但差异无显著性意义[(69.8±6.3)%,(92.4±4.2)%,(106.6±3.9)%,P>0.05]。提示LipofectamineLTX是人脐带间充质干细胞较为理想的化学转染试剂。

BACKGROUND：Genetic modification technology of human umbilical cord-derived mesenchymal stem cells（hUCMSCs） is numerous and varies in the complexity of their efficiency,reliability and safety.OBJECTIVE：To compare transfected efficiency of different novel chemical transfection reagents on hUCMSCs.METHODS：The hUCMSCs were transfected with Fugene HD,Lipofectamine LTX,and Attractene.Positive cells were detected under fluorescent microscope,and expression efficiencies were measured by using flow cytometry.Cell viability was measured by using trypan blue exclusion after transfection.RESULTS AND CONCLUSION：The highest transfected efficiency was observed with Lipofectamine LTX（32.5±2.12）%（P〈0.05）,followed by Fugene HD（4.3±0.64）%,minimal transfection efficiency was observed with Attractene（1.74±0.08）%.Lipofectamine LTX demonstrated the lower cell viability（69.8±6.3）% compared with Fugene HD（92.4±4.2）% and Attractene（106.6±3.9）%（P〈0.05）,but the difference was not significant.In summary,this study demonstrates that Lipofectamine LTX is a relative ideal chemical transfection reagent for hUCMSCs.