Title: Transfection efficiency of adenoviral vector expressing human hepatocyte growth factor with rat meningeal mesothelial cells in vitro and its effect on proliferation and migration
Abstract: Objective To investigate the transfection efficiency of adenoviral vector expressing human hepatocyte growth factor(Ad-HGF)with rat meningeal mesothelial cells(RMMCs)in vitro and its effect on proliferation and migration.Methods RMMCs were primary cultured and identified in vitro.Transfection efficiency was detected by fluorescence microscope and flow cytometer.The expression of HGF was measured with enzyme linked immunosorbent assay(ELISA)at different MOI of Ad-HGF or time points.The effect on proliferation of HGF was evaluated with MTT assay.The effect on migration of HGF was detected with Scratch wounds experiment and Transwells assay.All data were indicated by mean ± standard deviation and put to SPSS 13.0 sofaware.The linear regression equation was made through curve fitting analysis,t-test was applied to compare the data of two groups,and one-way ANOVA analysis was used to compare the data of groups.The standard test level was α=0.05.Results RMMCs were successfully isolated,cultureexpanded and identified in vitro,best MOI of adenovirus5 vector encoding enhanced GFP gene(Ad5-EGFP)was 200 that the transfection efficiency was 96.21%.The expression of HGF in the supernatant was increasing along with the increasing of MOI in a dose-dependent manner(F=407.851,P0.01),the highest concentration was(97.275±3.035)ng/ ml at 200 pfu/ cell.The expression of HGF was in a timedependent manner(F=384.063,P0.01),which became the peak of(104.275±2.356)ng/ ml after 48 h and sharply decreased on the 10th day then sustained at a stable level.The supernatant promoted the effect of proliferation in a does-dependent manner(F=285.946,P0.01).The supernatant promoted the effect of migration compared with control group(t=34.135,P0.01).Conclusions Ad-HGF can transfect efficiently RMMCs and the transfected cells can express the aim protein efficiently.HGF can promote the effects of proliferation and migration of RMMCs.
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
Access and Citation
AI Researcher Chatbot
Get quick answers to your questions about the article from our AI researcher chatbot