The Comparision Of The Function About Adipose Derived Stem Cells And Bone Marrow Mesenchymal Stem Cells In The Interverbral Disc Degeneration

Objectives:To separate,amplify and identify ADSCs and BMSCs；establish the model ofIntervertebral Disc Degeneration for the rabbit; To watch whether the transplanted stem cells cansurvive and proliferate in the degenerative disc,and discuss the two kinds of mesenchymal stemcells's function to the Degenerative DiscMeans:①The adipose tissue from rabbit was digested by collagenase and the primary adiposestem cslls were isolated from the adipose tissue.The rabbit bone marrow mesenchymal stem cellswere isolated and purified with entire bone marrow and attachment-independent culturesystem.A flow cytomter test the surface mark of ADSCs and BMSCs.The proliferation of cellswas observed by MTT method.②To establish the model of Intervertebral Disc Degeneration forthe rabitt by aspirating nucleus pulposus.③Transplant ADSCs and BMSCs marked by BrdU inthe Degenerative Disc of rabbit,after the operation and MRI inspection for 2,4,8weeks,thenexecute the experimental animals,and draw HE dyed material to survey the repairing situation inthe Degenerative Disc; Survey the survival and proliferation situation of cells dyed by BrdU inthe Degenerative Disc,Spectrophotometric determination with phlorglucinol test the changingsituation of proteoglycan content。Results:①ADSCs and BMSCs of the rabbit can be separated and purified in vitro,,proliferaterapidly,and have access to enough seed cells；FCM detects that the surface antihelion of ADSCsand BMSCs is CD29 positive,CD45 negative；MTT detects that the multiplication capacity ofADSCs and BMSCs is strong,the third generation celldigested and passaged transits intoplatform stage after incubation and logarithm growth period。②Aspirating nucleus pulposus canestablish the model of Intervertebral Disc Degeneration successfully.③2,4 and 8weeks after theoperation and MRI inspection showed that 3 groups intervertebral discs all regressed,and thegap narrowed down,signal of T2 weighting phase steped down.But Intervertebral DiscDegeneration of ADSCs transplant groups and BMSCs transplant groups is less serious thandefective regression group.④The transplanted ADSCs and BMSCs continued to exist in thedamaged intervertebral disc,and the cell is marked by 5-BrdU. The quantity of positive cell isbig in the second week, but less in the eighth week.With the time going , positive cell move tothe external area.⑤HE dying showed that ADSCs and BMSCs can survive and proliferate in theIntervertebral Disc Degeneration, and can also maintain in the organization structure of theintervertebral disc.⑥The quantity of proteoglycan ADSCs transplant groups and BMSCstransplant groups is more than that in the damaged intervertebral disc, showing significantdifference(P＜0.05).Results:①Collagenase digestion and entire bone marrow culture system can obtainmesenchymal stem cell in fat and marrow；②Aspirating nucleus pulposus will induceintervertebral disc Degeneration；③ADSCs and BMSCs transplanted to the intervertebral disc ofrabbit can survive and have multiplication capacity,can postpone the degeneration ofintervertebral disc and encourage the repair of intervertebral disc.In the histiocyte biological engineering of intervertebral disc degeneration, ADSCs and BMSCs can be both seed cell;Butthe success rate of separation ADSCs from marrow is higher than mesenchymal stem cell fromthe marrow, ADSCs is possible to replace BMSCs.