Empirical Study On The Survival And Osteogenic Differentiation Of EGFP-BMSCs In Vivo Of Host During Allograft Transplantation And Immunological

Objective:To observe the relationship between immunological rejection and osteogenic effect and quantitative change of heterogenic mice bone mesenchymal stem cells,BMSCs in vivo.Methods:1.The isolate,culture and purity appreciation of EGFP-BMSCs:bone marrow cells of EGFP-mice inoculated in the culture bottle to cultivate.By way of changing the culrure medium gradually,the cells which didn't stick to the wall may being taken away and EGFP-BMSCs obtain purification.and detect the purity of EGFP-BMSCs by flow cytometry when fifth passage.2,To digest the EGFP-BMSCS from the same mice.and become the cells density is 2×10~6/ml.we injected about0.1 ml EGFP-BMSCs into the right tibia muscle of each BALB/C mices as allograft model.The same muscle on the left leg was injected 0.1ml isotonic Na chloride as muscle control group.Then to observe the appearance of Injection site.3,The flow cytometry was used to analysis the changes in the ratio of T lymphocyte subsets of CD4+,CD8+ in the parasitifer's peripheral blood at different times.The legs were cut to Frozen section in order to make sure that if there were enough EGFP-BMSCs to be observed by fluorescence microscope at differente experimental times.And then,aim to explore the Inflammatory reaction and bone formation.The slices were identified by HE staining and immuno-histochemistry staining of Collagen-I. Results:1,Apply the method of different speed of adherence,change culture medium and passage to isolate and culture EGFP-BMSCs can obtain enough quantity of EGFP-BMSCs.2.After allograft,EGFP-BMSCs,there were not obviously abnormal reaction at parasitifer,and the colour of injection site were short of change.3.After allograft transplantation of EGFP-BMSCs,the EGFP-BMSCs can be survived about one week.After allograft,EGFP-BMSCs didn't show the characteristic of into osteoblast at different experimental times.4.There were not inflammatory cell in the muscle of mice legs at different experimental times5.Fluorescence microscope observation of mice legs make know that EGFP-BMSCs was detected in one day and one week.and display the amount of allograft EGFP-BMSCs play a reduced tendency role.6,The peripheral blood T lymphocyte subsets CD4+,CD8+ ratio of different experiment groups had no significant difference compared to the empty contrast group.Conclusion:Host did't have acute or conspicuous rejection performance after allograft transplantation of EGFP-BMSCs.The allogene EGFP-BMSCs didn't show the characteristic of into osteoblast.