Study of BMP2 and EGFP recombinant adenovirus transfection on rabbit bone marrow derived mesenchymal stem cells in vitro
10.3969/j.issn.1671-8348.2014.02.023
- VernacularTitle:BM P-2和EGFP重组腺病毒体外转染兔骨髓间充质干细胞的研究
- Author:
Jiabin CHEN
;
Qiang LI
;
Jia RU
;
Chengcong WU
;
Yingkuan NING
;
Weiliang CAI
- Publication Type:Journal Article
- Keywords:
bone morphogenetic proteins;
adenoviridae;
bone marrow;
mesenchymal stem cells;
gene;
transfection
- From:
Chongqing Medicine
2014;(2):193-195,199
- CountryChina
- Language:Chinese
-
Abstract:
Objective To culture the rabbit bone marrow derived mesenchymal stem cells (BMSCs) ,and to observe their biologi-cal characteristics in vitro and the expression of BMP2 and EGFP after Ad-EGFP-BMP2 infected on them .Methods To isolate and cultivate BMSCs by density gradient centrifugation and adherent culture in vitro ;the cell cycle and the surface marks were detected by flow cytometer ;rabbit BMSCs were differentiated into the direction of osteogenetic cells by in vitro induction .After transfection , the expression of exogenous gene in the cells was detected by the immunocytochemical staining and Western blot .Results About 56 .84% of rabbit BMSCs cell cycle was in the G1 phase;the D44 expression was positive and the CD45 expression was negative ;af-ter induction by osteogenetic cells ,the Ⅰtype collagen immunohistochemical staining was positive and the alizarin red staining was positive .After transfection ,the strong expression of BMP2 and EGFP in cells was showed by Western blot and the immunocyto-chemical staining .Conclusion rabbit BMSCs are successfully isolated ,cultured and identified to have the ability of osteogenetic dif-ferentiation ;the structured Ad-BMP-2/EGFP can efficiently transfected rabit BMSCs and stably express the target gene of BMP 2 .
