BMSCs Induced Into Nucleus Pulposus-like Cells In Vitro Studies

[Objective]1.Isolation, expansion and identification of Rabbit BMSCs；2.BMSCs was induced into nucleus pulposus-like cells，then find the right time for thecell transplantation.[Methods]Application pure stick wall culture method isolation, expansion rabbit BMSCs andnucleus pulposus cells，and use immunocytochemistry identify BMSCs surfacerelative specific molecular(CD90、 CD44、 CD45、CD34) to determine thepurification of BMSCs. Transwell chamber which contains BMSCs was placed in cellculture dish which contains the nucleus pulposus cells，establish the co-culturemodel，through the cell vitality、RT-PCR（reverse transcription PCR）and Westernblot detect collagen type II and aggrecan，find the best co-cultured time in vitro toprovide more experimental data with disc tissue engineering of seed cells furtherstudy.[Results]BMSCs and nucleus pulposus cells were successfully isolated， expanded, andimmunocytochemistry showed that the positive rates of specific molecular surfacemarkers of BMSCs were93.2%of CD90，95.5%of CD44，4.4%of CD45，5.5%ofCD34. Through the cell viability、RT-PCR and Western blot detected,we find thatBMSCs can differentiated nucleus pulposus-like cells in vitro,and achieve the bestcondition about15days,the cells can appropriate in vivo transplantation.[Conclusions]1.The pure stick wall of separation can successfully isolate the high purity of BMSCs.2. Transwell chamber and cell culture dish constitute a culture system，and BMSCscan differentiated nucleus pulposus-like cells in this co-cultured system.BMSCs candifferentiated nucleus pulposus-like cells and achieve the best condition about15days,and at this time the cells can appropriate in vivo transplantation.