Transfection of Lentivirus Recombined with Marker Gene into Human Umbilical Cord Wharton’s Jelly-Derived Mesenchymal Stem Cells
10.3969/j.issn.0253-9896.2013.10.012
- VernacularTitle:携带标记基因的慢病毒载体转染人脐带华通胶间充质干细胞的实验研究
- Author:
Li WANG
;
Xiaohong XU
;
Ningkun ZHANG
;
Lianru GAO
;
Zhiming ZHU
- Publication Type:Journal Article
- Keywords:
mesenchymal stem cells;
flow cytometry;
lentivirus;
genetic vectors;
gene therapy;
transfection;
cell pro-liferation
- From:
Tianjin Medical Journal
2013;(10):985-988,1044
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the optimal condition of lentivirus,which was recombined with marker gene of enhanced green fluorescent protein (Lentivirus-EGFP) transfect human umbilical cord wharton’s jelly-derived mesenchy-mal stem cells (HUWMSCs) and the effect of transfection on the proliferation in HUWMSCs. Methods HUWMSCs were transfected with EGFP by lentivirus vector in vitro via different multiplicity of transfection (MOI) in four different transfec-tion methods (A, B, C and D). The fluorescence expression and the transfection efficiency in different methods were analyzed by both fluorescent microscope and flow cytometry. The proliferation rates of infected HUWMSCs was evaluated by MTT method. Results The transfection efficiency was 10.6%-87.3%after 4 days in all experimental groups, which showed the dose-effect relationship with MOI. Polybrene (5 mg/L) could significantly increase the transfection efficiency (P＜0.05). Re-sults of MTT assay showed that there were significant differences in the proliferation rates of infected HUWMSCs between different transfection methods (P ＜ 0.001). There was better cell proliferation in method A (MOI=10) group and method B (MOI=10) group than that of other groups. Conclusion Method B (MOI=10) is the optimal transfection method in this exper-iment. HUWMSCs could be transfected by lentivirus-EGFP with high efficiency and could stably express transfected gene within 2 weeks, which is a safe and effective gene transfer vector.
