Rabbit Adipose Mesenchymal Stem Cells And Nucleus Pulposus Cells Between Trained Again After Transplantation Of Intervertebral Disc Degeneration

Objective To study the reimplantation of co-cultured adipose-derived mesenchymal stem cells and nucleus pulposus cells of intervertebral disc(IVD), and observe its effect on type collagen and proteoglycan in rabbits.Methods (1)To isolate and culture adipose-derived mesenchymal stem cells and nucleus pulposus cells of rabbits in vitro. (2) To verify the multipotential differentiation of mesenchymal characteristics of ADSCs, the ADSCs were subjected to differentiation in conditions known to induce adipogenic and osteogenic lineages.(3) Let ADSCs and NPCs culture together to make ADSCs differentiate into NPCs. (4) 30 New Zealand white rabbits were devided into experimental group and degeneration model control group, to choose L3/4、L4/5、L5/6 IVD of rabbits as experimental discs,the aspirated IVDs of experimental group were injected with cell suspension of ADSCs and NPCs, but the oher group were only injected with PBS. The IVDs of the two groups were taken out 4 weeks and 8 weeks later. The changing situation of proteoglycan content tested by spectrophotometric determination with phlorglucinol,gene expressions of type II collagen and proteoglycan were detected by reverse transcription polymerase chain reaction(RT-PCR) and analyzed bysemiquantitative technique.Result ADSCs can be induced to differentiate into adipogenic and osteogenic lineages. Spinedle or multi-angular ADSCs turned into fibro-like phenotype in the co-culture with NPCs, and the expressions of typeⅡcollagen and proteoglycan in degeneration model control group were weaker than that in experimental group(P<0.05).Conclusion Collagenase digestion can obtain mesenchymal stem cell in fat of rabbit. Retransplantation of co-cultured adipose-derived mesenchymal stem cells and nucleus pulposus cells in rabbits can delay the degeneration of the intervertebral discs.