Primary Study On The Regular Of Survived And Differentiated Into Chondrogenic About The BMSCs In Host's Muscle After Allograft

【Objectives】:The rapid evolution of experimental data has acknowledged the impotance of immune biology,survived and potential differentiated characteristics of bMSCs.So it is necessary to estimate the immune property of bone marrow mesenchymal cells,and explore the survived and differentiated rules of MSCs in vivo after allograft.All the results of this experiment can supply more detailed theoretic dates about the basic cells of bone tissue engineering.【Method】:1.To establish a successful system for the separation,culture and identification of MSCs from GFP(green fluorescent protein,GFP)gene transinfected FVBN mouse in vitro.2.36 BALB/C mouse were divided into 6 groups randomly.We injected 2×10~5 GFP transinfected MSCs to the right tibia muscle of each BALB/C mouse as allograft model,The same muscle on the left leg was injected 0.1ml distilled water as control group.Executing mouse at the time before injecting,and ldays,1,2,3,5,7weeks after injecting.3.(1) The flow cytometry was used to analysis the changes in the ratio of CD4+CD25+(T regular cell) in the host's Peripheral blood at different experimental times.(2) In order to explore the survived and differentiated regular of allograft MSCs in vivo,right leg muscle of host was cut to frozen sections,fluorescent stereoscope to visualize and record the fate and the location of grafted MSCs at different experimental times,then the slices were evaluated by immuno-histochemistry staining of CollagenⅡantibody to explore weather the MSCs can differentiate to chondrogenic cells.【Results】:1.Bone marrow mesenchymal stem cells which were transinfected by GFP gene were isolated and cultivated successfully.2.The T regular cells in peripheral blood of host between different experimental time groups had no significant difference compared to the empty contrast group(P) in all the stages.3.The bMSCs can be observed in host's muscle in one week after allograft,then they can not be found after that.They had no characteristics of chondrogenic differentiation in vivo.【Conclusion】:1 The combination of all bone marrow culture and adhere culture is an effective method to isolate and culture MSCs.2.The little changes of T regular cells in peripheral blood of host suggest the rejection of allograft was significant low,which indicate low immune property of the bone marrow mesenchymal stem cells as engraftment stem cells.3.The MSCs can be observed in host's muscle inner one week after allograft,and the MSCs didn't show the characteristics of differentiating into chondrogenic cells.This may be because of the cells died,migrated to other part in vivo or this microenvironment can not induce the MSCs to chondrogenic cells.