Study On Bone Marrow Mesenchymal Stem Cells And Adipose Derived Mesenchymal Stem Cells In Osteogenic Differentiation Under Different Proportion Of Mixed Culture

Objective : The purpose of this experiment is to study the bone marrow mesenchymal stem cells(BMSC)and adipose derived mesenchymal stem cells(ADSC)co culture influence on stem cell osteogenic ability in different proportion.Method:By using adherent separation and purification were obtained from adipose derived mesenchymal stem rats method and enzyme digestion method Cells and bone marrow mesenchymal stem cells.In vitro culture environment of the two kinds of stem cells into chondrocytes,respectively,osteogenic differentiation of adipose tissue into.On chondrogenic induction group were identified;bone differentiation group by alizarin red staining,while fat group,oil red O staining with toluidine blue reagent.After osteogenic induction group were divided into 6 groups.Group A: to induce the undifferentiated stem cells in the blank control group;group B:bone marrow mesenchymal stem cells in the osteogenic induction group;group C adipose derived mesenchymal stem cells osteogenic induction group;group D: BMSCs and ADSCs mixed induced group,ADSC and BMSC cell proportion is 1:3;Group E:BMSC and ADSC proportion is 1:1 mixture induced group;Group F: BMSC and ADSC ratio is 3:1 mixed in the induced group.The six groups were taken the cell density of1.0×104/cm2 planting.The above six groups of cells were detected by Western-Blot RUNX2 and OCN protein.In vivo experiment of randomly selected different sex Lewis thirty-five rats were divided into experimental group and control group,all the rats are made of skull defect model.The control group was implanted biological scaffold material blank,the first experimental group six group cells into biological scaffolds,then implanted in the skull defect.Six weeks later the material removed alizarin red staining of calcified foci,and by Dr chip shooter on the rat skull recovery were analyzed.RESULT:ADSC and BMSC cells in chondrogenic cells induced into cartilage differentiation,toluidine blue staining visible blue;osteogenesis induced osteogenic differentiation of cells under differentiation conditions,alizarin red staining showed a patchy red;adipogenic conditions induced into adipocytes and oil red O staining visible red in color.In osteogenic differentiation of six groups.BCDEF cells in each group of osteocalcin and Runx2 protein expression level was significantly higher than that of cells in group A(P < 0.05);group B cell protein expression was higher than that of C group(P < 0.05);in def cells protein expression than that in B and C group increased significantly(P < 0.01);def in three groups,the protein expression of volume of the order followed by F>E>D.In vivo experiments for six weeks later will implant materials and skull after removing found: alizarin red staining of the experimental group had calcification foci formation and the amount of calcification for mixed group > alone induced group > in the non induced group,Dr shot out of the poor(see behind the analysis discussion).Conclusion: After Co cultured osteogenic ability of two kinds of cells increased.As scaffold material for bone cells were implanted into the bone defect model in the rat in vivo can promote bone healing and mixed cells induced by osteogenic ability is alone induced strong and mixed culture of bone marrow mesenchymal stem cells in bone is composed of ability better.