The Expression Of Green Fluocescent Protein Gene Transfected The Bone Marrow Stromal Cells With Lipofectin

Objective Bone marrow stromal cells are considered as an ideal sources of seed cells with the advantage of being isolated and proliferated in vitro easily, which is a prerequisite for the autotransplantation.If the cell could be easily accept and express the exogenous gene,it would be more widely used in the furture study. Therefor the objects of the present study are:1 To establish an effective method for isolation and culturation of the rats' bone marrow stromal stem cells.2.To establish the possibility of the expression of green fluocescentprotein gene transfected the bone marrow stromal cells with lipofectin.Methods After isolated by density gradient contrifugation with Percoll,the rats' bone marrow stromal stem cells were cultured with DMEM culture medium containing 20% fetal bovine serum in the plastic flastic. When the cells covered the flask, they were passaged with pancreatin. Their morphology were observed under inverted microscope. Green fluocescent protein gene was transfected with lipofectin the bone marrow stromal cells that were isolated from Wister rats and explanded in vitro. Then the cells were observed under a fluorescence microscope to examine the gene expression. And the cell vitality was detected. Results1. The vitality of the isolated cells was 99%.After cultured in the plastic flask for 10-14 days, the cells covered the bottom and were passaged. Then they werepassaded every 5-7 days. The typical morphology was seen after the cells were passaged for 3-4 times.2. The green fluocescent protein gene expressed in the bone marrow stromal cells with an transfection efficiency of 28.9% ±3.6%. The cell vitality has no significant difference between the experiment and control group.times.Conclusion Rats' bone marrow stromal cells can be isolated by density gradient contrifugation with Percoll and can be gene-modified with lipofectin .