Human Bone Marrow Derived Mesenchymal Stem Cells Proliferation Through Beta-catenin/Wnt Pathway Activation

Objective: Explore the function of beta-catenin/Wnt pathway in the regulation of marrow-derived mesenchymal stem cells proliferation.Methods: First isolate and culture the bone marrow derived MSCs,identify MSCs surface markers,and test the multipotential differentiation capacity of MSCs.And use lithium,GSK3? inhibitor SB216763,quercetin and si RNA mediated knock down of beta-catenin,to upgrade or downgrade beta-catenin/Wnt signal pathway,and then determine the proliferation rate of cells with CCK-8.Results:After isolate and culture,hematopoietic,endothelial,and myeloid cell lineage-specific antigens,such as CD34,CD133,CD45,CD146,and HLA-DR were not expressed in these cells.Most of the cells were positive for CD29(95±3.7%),CD44(91±2.1%),CD73(98±1.4%),and CD90(98±2.3%)antigen.Approximately 76±6.2% cells expressed CD105.These cells can be readily differentiated into adipocyte,osteocyte,and chondrocyte cells by culturing in appropriate induction media,as determined by corresponding staining.Li Cl increased MSCs in a dose depended manner and toped at 5m M.At this concentration,Li Cl increased OD value by proximately 1.4 fold versus control groups.SB216763,a specific GSK-3? inhibitor,effect on MSCs proliferation similar to lithium,SB216763 increased MSCs total cell number by approximately 1.6 fold versus control,which toped at 50?M.We then treated with 5m M Li,and a series concentration of Que for 6 days.CCK-8 assay showed that quercetin apparently abolished the proliferating effect of Li at concentration of 30?M.In ?-catenin knocked down MSCs,CCK-8 assay after 6 days revealed that lithium was deficient in their ability to stimulate proliferate.Conclusion:Beta-catenin/Wnt pathway played an important role in on the regulation of human mesenchymal stem cells proliferation.Regulate beta-catenin/Wnt pathway by small molecule drugs can effectively influence proliferation of MSCs.This founding can provides a theoretical basis to the treatment of beta-catenin/Wnt pathways related diseases disorders of bone metabolism.