Osteogenic induction of rabbit's bone marrow stromal cells and transfection of AAV-EGFP in vitro
- VernacularTitle:兔骨髓基质干细胞成骨诱导及增强型绿色荧光蛋白基因腺相关病毒体外转染的研究
- Author:
Zhao-Min ZHENG
;
Zhi-Yong DONG
;
Guan-Ming KUANG
;
Fobao LI
;
- Publication Type:Journal Article
- Keywords:
Bone marrow stromal cells(BMSCs);
Osteogenic induction;
Adeno associated virus (AAV);
Enhanced green fluorescent protein (EGFP);
Transfection
- From:
Chinese Journal of Orthopaedic Trauma
2004;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the effects of adeno-associated virus-enhanced green fluorescent protein (AAV-EGFP)on the biologic behavior of rabbit's bone marrow stromal cells(BMSCs)by means of a simple method of culturing and osteogenic induction in vitro so as to find an ideal viral vector and cell tracing mark for tissue en- gineering.Methods Total bone marrow culture was conducted to obtain rabbit BMSCs which were then induced in the osteogenic direction.The morphology of the cells was observed continuously,and their surface antigen and ossification were detected by alkali phosphatase stain and Von Kossa stain.On the basis of the above results, AAV-EGFP was transfected into the induced cells.The morphologic changes of the cells,the expression time and intensity of fluorescent light were observed.The transfection efficiency was detected to find the best multiplicity of infection(MOI)value.The cell growth curves were drawn to evaluate the biologic effects of AAV-EGFP on the cyto-activity.Results The morphology and purity of the rabbit BMSCs obtained were good.The ossification of the cells was significant after osteogenic induction.The best MOI value was found to be 1?10~5.The expression intensity of fluorescent light was strong with the expression time more than eight weeks so that the fluorescent light could be observed after cell generations.The transfection efficiency of AAV was high without significant biologic effects on the cyto-activity.Conclusions The total bone marrow culture and in vitro cell induction can satisfy the requirements for seeding cells in tissue engineering.AAV is an ideal viral vector for tissue engineering.Transfection of AAV-EGFP to cells could be an ideal method for cell tracing mark.
