[Efficiency of human and rat bone marrow mesenchymal stem cells transfected by modified adenovirus].
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).


CONCLUSIONS
AdF35 may be the ideal vector to carry the target gene to transfect hBMSCs effectively but not to transfect rBMSCs.
Hu
Shuo
S
Department of Nuclear Medicine, Xiangya Hospital, Central South University, Changsha,China. hushuo_xy@sina.com
Xiong
Lingjing
L
Yu
Jianying
J
Lei
Meng
M
Zhao
Min
M
Zhang
Yongxue
Y
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D014162:Transfection
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins