The Effect Of Chondrogenic Pre-differentiation Of Adipose-derived Mesenchymal Stem Cells For Regeneration In Harsh Intervertebral Disc Microenvironment

ObjectiveRecent studies indicated the cell based therapy could be a promising approach to treat intervertebral disc(IVD)degeneration.Though the harsh microenvironment in disc is still challenging to implanted cells,it could be overcome by pre-conditioning graft cells before transplantation,suggested by previous literatures.Therefore we designed this study to identify the potential effect of chondrogenic pre-differentiation on adipose-derived mesenchymal stem cells(ADSCs)in IVD-like microenvironment,characterized by limited nutrition,acidic and high osmosis in vitro.MethodsADSCs of rat were divided into five groups,embedded in type II collagen scaffold and cultured in chondrogenic differentiation medium for 0,3,7,10,14 days respectively.Then the ADSCs were implanted and cultured in IVD-like condition.The proliferation and differentiation of ADSCs was evaluated by cell counting kit-8 test(CCK-8),Real-time quantitative polymerase chain reaction(RT-PCR),and Western blotting and immunofluorescence analysis.ResultsAnalyzed by the first week in IVD-like condition,the results showed relatively greater proliferative capability and extra-cellular matrix(ECM)synthesis ability of the ADSCs pre-differentiated for 7 days and 10 days than the control.ConclusionsWe concluded that pre-differentiation of rat ADSCs in chondrogenic culture medium for 7-10 days could promote the regeneration effect of ADSCs in IVD-like condition,and the pre-differentiated cells could be a promising cell source for disc regeneration medicine.