| Background:Repair of bone defects is an arduous task in plastic and reconstructive field.Autologous bone transplantation was considered as the golden standard implants.But for its limitations,this could not be used widely for bone defects repair.The boost of bone regenerative medicine seems to provide a new solution for this: lab making bone grafts with autologous stem cells and scaffolds.Nevertheless,there are three critical barriers need to be crossed: enough seeding cells,less inmuno-reactive scaffold and relative growth factors.Bone marrow derived stem cells(BM-MSCs)are considered as an ideal seeding cells for its osteogenic potential.To get enough cell number,the in-vitro culture usually was used to expansion.But,the multilineage potential of BM-MSCs also was weakened during the expansion.And if the bone graft were larger than usual,BM-MSCs can’t provide sufficient seeding cells.Adipose derived stem cells might replace BM-MSCs for its high and multilineages potential including osteogenesis.Meanwhile,extracellular matrix(ECM)could contribute to the adhesion,proliferation and differentiation of stem cells cultured on it.Objects:In this study,we tried to explore effects of human bone marrow stem cells derived extracellular matrix on the adhesion,proliferation and differentiation of human adipose derived stem cells.Methods:Initial human ASCs were seeded at same seeding density on conventional plastic culture dishes and dishes coated with hBM-ECM.Cells at 2 passages were collected for further research.The proliferation and the potential of osteogenesis and adipogenesis of h ASC expanded from hBM-ECM were determined by MTT,CFU-Fs,-AD,-OB(colony forming units-fibroblast like colonies,-adipogenesis,-osteogenesis).Real-time PCR was also used to determine the relevant gene expression.After culture expansion,h ASC were transplanted in nude mice to assess the in-vivo osteogenesis.Results:After 2 passages expansion of initial cells,hBM-ECM could provide about 3.27 folds more cells than conventional culture dishes.There were no proliferation and self-renew advantages once they left hBM-ECM.Cells could form more adipogenic colonies and fewer osteogenic colonies after 2 passages expansion on hBM-ECM comparing with cells expanded on conventional culture dishes.Osteogenic relevant gene expression was increased in the cells expanded on hBMECM than cells on plastic dishes.The in-vivo osteogenesis analysis also showed cells cultured on hBM-ECM could generate more bone collagen.Conclusions:Compared with conventional culture dishes,hBM-ECM could improve hASCs proliferation and enhance its osteogenic potential but inhibit its adipogenesis potential.According the results,hBM-ECM is an ideal expansion system for hASCs in bone regenerative medicine. |
