Efficiency of human and rat bone marrow mesenchymal stem cells transfected by modified adenovirus.
10.3969/j.issn.1672-7347.2010.09.014
- Author:
Shuo HU
1
;
Lingjing XIONG
;
Jianying YU
;
Meng LEI
;
Min ZHAO
;
Yongxue ZHANG
Author Information
1. Department of Nuclear Medicine, Xiangya Hospital, Central South University, Changsha,China. hushuo_xy@sina.com
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
Animals;
Bone Marrow Cells;
cytology;
metabolism;
Genetic Vectors;
Green Fluorescent Proteins;
biosynthesis;
genetics;
Humans;
Mesenchymal Stem Cells;
cytology;
metabolism;
Rats;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2010;35(9):983-989
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To compare the efficiency of human bone marrow mesenchymal stem cells (hBMSCs) with rat BMSCs (rBMSCs) transfected by modified adenovirus containing fiber 35 (AdF35)-enhanced green fluorescence protein(eGFP).
METHODS:We separated hBMSCs and rBMSCs from the bone marrow of humans and rats, respectively, and osteogenesis and adipogenesis were induced. eGFP was carried by modified AdF35, which was transfected to hBMSCs and rBMSCs with different multiplicity of infections (MOIs). Activity of the cells was detected by MTT. The transfected cells were observed under fluorescent microscope. The transfection efficiency was measured by flow cytometer. The expression of coxsackie and adenovirus receptor (CAR) and CD46 mRNA in the cells was inspected by real time PCR.
RESULTS:hBMSCs and rBMSCs induced osteogenesis and adipogenesis successfully after being separated from human and rat bone marrow respectively. The activity of the cells was inhibited when MOI was 1,000 PFU/mL. hBMSCs with strong green fluorescence were observed but few rBMSCs were seen under fluorescence microscope 48 h after being transfected by AdF35-eGFP. The transfective efficiency was (84.8±7.1)% and (3.3±1.1)%, respectively. The expression of CD46 was high while that of CAR was low in hBMSCs. The expression of CAR was very high and that of CD46 was low in rBMSCs (P<0.01).
CONCLUSION:AdF35 may be the ideal vector to carry the target gene to transfect hBMSCs effectively but not to transfect rBMSCs.
