[Study of infective efficiency of two adenovirus vectors to rBMSC and its mechanism].
OBJECTIVE
To investigate the infective efficiency of Ad5/EGFP versus AdF35/EGFP to rat bone marrow mesenchymal stem cells (rBMSC) and understand its mechanism.


METHODS
rBMSC were isolated from rat bone marrow by density gradient method. The expressions of CD90, CD29, CD44, CD34, CD11b and CD45 on the surface of rBMSC were measured by flow cytometry. The EGFP-carrying Ad5 and AdF35 were infected into rBMSC respectively. The effect of Ad5/EGFP and AdF35/EGFP with different MOIs to rBMSC was tested by MTT. Infected rBMSC were observed by fluorescence microscopy. The infective efficiency and mean fluorescence intensity were determined by flow cytometry after a cellular infection for 1 - 9 days. The expressions of CAR and CD46 on rBMSC were measured by real-time PCR.


RESULTS
The expressions of CD90, CD29 and CD44 on rBMSC were positive while those of CD34, CD11b and CD45 were negative. When MOI </= 1600 and </= 1000, Ad5/EGFP and AdF35/EGFP could not inhibit the growth of rBMSC respectively (t = 3.12, P = 0.052; t = 2.20, P = 0.15). RBMSC with EGFP expression could be observed under fluorescence microscope after a 1-day infection by Ad5/EGFP. The number of observed cells peaked after an infection for 2-3 days and then decreased gradually. And a few cells could be still observed for 9 days. Fewer cells could be observed after an infection by AdF35/EGFP for 1 - 9 days. The infective efficiency of Ad5/EGFP was significantly higher than that of AdF35/EGFP (t = 2.45, P = 0.008). There was a high expression of CAR on rBMSC. The expression of CD46 was quite low. There was significant difference between them (t = 2.57, P < 0.01).


CONCLUSIONS
The infective efficiency of Ad5/EGFP is significantly higher than that of AdF35/EGFP. It may explain the expression patterns of CAR and CD46 on rBMSC. AdF35 is not a suitable gene-carrying vector to infect rBMSC. Yet Ad5 can be an efficient vector carrying target gene to infect rBMSC for curing diseases.
Hu
Shuo
S
Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, China.
Xiong
Ling-jing
LJ
Yu
Jian-ying
JY
Lan
Xiao-li
XL
Zhang
Yong-xue
YX
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001854:Bone Marrow Cells; D002460:Cell Line; D005434:Flow Cytometry; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D051381:Rats; D017207:Rats, Sprague-Dawley; D011991:Receptors, Virus / Q000235:genetics*; D014161:Transduction, Genetic; D014162:Transfection
D011991:Receptors, Virus