GDF-5 And Rabbit Disc In Vitro Model Induction BMSCs Of Rabbit Differentiation To The Nucleus Pulposus Cell

Objective to explore the GDF-5 and rabbit disc in vitro model induction BMSCs of rabbit differentiation to the feasibility of nucleus pulposus cell, for the next GDF-5 rabbit BMSCs and carrying scaffold materials into living rabbit disc model lays experimental basis.Methods using all the marrow cells stick wall separation cultivate rabbit mesenchymal stem cells, inverted microscope observe BMSCs form, CD54 CD44 immunohistochemical stains and identification, PI dyeing measuring the cell cycle, CCK-8 detection P3,P5,P7 cell proliferation, detection P3,P5,P7 cell stick wall rate, RT-PCR detection cell type ii collagen, the proteoglycans, MMP1, MMP2, ADAMTs-5 gene expression. Media, including ordinary medium, containing 100ng/mlGDF-5 medium, containing 100ng/mlGDF-5 medium combined with rabbit intervertebral disc in vitro model, respectively cultivate mesenchymal stem cells. PI dyeing measuring the cell cycle.CCK-8 detection cell proliferation. Immunohistochemical, alcanblue dyeing, toluidine blue dye detection type ii collagen and proteoglycans expression.Results BMSCs form shuttle or spindle or polygonal, state clearly, surface antigen markers CD44, CD54 immunohistochemical stains positive,PI dyeing cell cycle show for slow cycle cells, P3,P5,P7 cells proliferation and stick wall rate attenuation trend. BMSCs don't express type ii collagen and proteoglycans genes, express MMP1, MMP2, ADAMTs-5 genes. After cultivate mesenchymal stem cells respectively, containing 100ng/mlGDF-5 medium combined with rabbit intervertebral disc in vitro model group cells form bigger to go round.Containing 100ng/mlGDF-5 medium obviously accelerate cell proliferation, S period count increase noticeable cell, typeâ…¡collagen immunohistochemical, alcanblue dyeing, toluidine blue staining: negative. Containing 100ng/mlGDF-5 medium combined with rabbit intervertebral disc in vitro model group cells beginning proliferation dropped significantly, S period cell count also increased, typeâ…¡collagen immunohistochemical, alcanblue dyeing, toluidine blue staining: positive.Conclusion GDF-5 with rabbit disc in vitro model induction can make BMSCs of rabbit differentiation to class nucleus pulposus cell, the rabbit BMSCs induced training with the function of secrete typeâ…¡collagen and of proteoglycans, provided the possibility for disc regeneration.