Impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells
10.3969/j.issn.1006-5725.2016.06.004
- VernacularTitle:腺病毒对原代枯否细胞转染效率及增殖活性的影响
- Author:
Tong ZHU
;
Wenjuan TU
;
Zhili TAN
;
Liangming LIU
- Publication Type:Journal Article
- Keywords:
Primary kupffer cells;
Adenovirus;
transfection;
Green fluorescent protein;
Cell proliferation
- From:
The Journal of Practical Medicine
2016;32(6):879-882
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells (KCs). Methods Rat liver KCs were separated and purified by density gradient centrifugation , and was then transfected with adenovirus carrying green fluorescence protein (GFP) gene at different multiplicity of infection (MOI). After 24 h, the transfection efficiency was evaluated by fluorescence microscope and flow cytometry. The proliferative activity of KCs was assessed by colorimetric method. Results The positive percentages of GFP staining cells were statistically different among different doses of adenovirus (MOI 0, 100, 300, 500, 700 and 900) under fluorescence microscopy or by flow cytometry (P <0.05 for all comparisons). The cell proliferative activity had significant differences among MOI 300, 500, 700 and 900(P < 0.05 for all comparisons), but had no differences among MOI 0, 100 and 300 (P > 0.05 for all comparisons) by CCK8 assay. Conclusions KCs can effectively be transfected by GFP adenovirus; and with an increase in virus MOI, the transfection efficiency rises gradually. A higher dose of adenovirus may have a negative effect on cell proliferative.
