Experimental Study Of Bone Marrow Stromal Stem Cells Differentiation Into Vascular Endothelial Cells

IntroductionVascularized and bone regeneration are both the most basic aspect in the bone healing process.The rapid development of bone tissue engineering provide the ideas in line with the principles of biology for bone regeneration.The research in the field focused on the access of Seed cells and the exploring for the conditions of inducation. marrow mesenchymal stem cells receive much attention because of their low immunity,differentiation in several directions and proliferative potention.Our experiments will explore the use of vascular endothelial growth factor(VEGF165) induced bone marrow-derived mesenchymal stem cells differentiation into vascular endothelial cells.So as to provide new seed cells for tissue engineering,exploring the experiment conditions at the same time.ObjectiveTo study differentiation of bone marrow-derived mesenchymal stem cells(BMSCs) into blood vessel endothelial cells for ideal cell origin of complex organ tissue engineering vascularization and injured tissue repairing by cell transplantation.To Discuss the effects of the different concentrations of vascular endothelial growth factor(VEGF₁₆₅)on bone marrow mesenchymal stem cell proliferation and differentiation into endothelial cells.Materials and Methods1.Materials(1)Specimen source:Wistar Rats(Weight about 100g,Three-week-old,Male and female open,Laboratory Animal Center of China Medical University Provide.(2)Major reagents:Vascular endothelial growth factor(VEGF165),Rabbit anti-mouse VEGFR1/Flt1 polyclonal antibody,rabbit anti-goat SABC kits2.Methods We use density gradient centrifugation and attaching growth to isolate and purify rat BMSCs.Group A is negative control group.5ng/ml,10ng/ml,50ng/ml,100 ng/ml VEGF₁₆₅were seperately added to group B,C,D and E.After 24 hours of culture,a comparison was made between four groups for cell proliferation.After 14 days of cultures,observations were made for morphological changes,The differentiated cells were identified by measuring surface marks(VEGFR1/Flt1)on the 14th day.ResultsFor the cultured bone marrow mesenchymal stem cells in vitro,Different groups of culture medium containing different concentrations of VEGF₁₆₅have different effects on bone marrow mesenchymal stem cell proliferation.There are significant differences.Experimental groups can induce bone marrow-derived mesenchymal stem cells differentiation into vascular endothelial cell compared with the control group.The proportion of positive cells Are separately 2.1598±0.2555%,30.3591±5.1687%,44.7643±2.1011%,56.0111±2.9603%,86.2438±0.9663%.There are also significant Differences between the experimental groups.ConclusionBone marrow mesenchymal stem cells have the ability of differentiation into vascular endothelial cells.They can be used as seed cells in tissue engineering. VEGF165 can promote bone marrow mesenchymal stem cell proliferation and differentiation into endothelial cells.And the level of the proliferation and differentiation into EC was correlated with the dose of VEGF165.