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Background:Recently,stemcelltherapyhasbeenattractinggreatinterestasapotentiallyeffectivetreatmentofdegenerativediskdiseases.Manystudiesindicatethatmesenchymalstemcells(MSCs)havethecapacitytorepair degenerativedisksbydifferentiatingtowardchondrocyte-likecells,whichproduceextracellularmatricesespeciallyaggrecanandtypeIIcollagen.However,themicroenvironmentwithinthedegenerativedisks,whichischaracterizedbyhypertonicity,acidityandlowglucose,appearstobechallengingtotheimplantedcells. Purpose:Thisstudywasaimedtodetectthedifferentresponsesoftheadipose-derivedmesenchymalstemcells(ADSCs)intheenvironmentmimickingdegenerativedisk,whichwerepre-processedfordifferenttimelengthsinchondrogenesisdifferentialculturemedium.Method:RatADSCswhichweredividedintofivegroupswereculturedrespectivelypre-processedfor0,3,7,10,14daysinchondrogenesisdifferentialculturemedium,thenculturedinthedegenerative-disk-likedifferentialculturemediumfortheresttime,3weeksintotal.Cellviabilitywasmeasuredbylactatedehydrogenase(LDH),cellproliferationbeingmeasuredby CellCountingKit-8(CCK-8).Theexpressionofaggrecan,typeIcollagen,andtypeIIcollagenwasdetectedbyreal-time quantitativepolymerasechainreaction,meanwhiletheexpressionatproteinlevelwasdetectedbywestern-blottinganalysis.Result:Thedegenerative-disk-likemicroenvironmentslightlyinhibitstheviabilityandproliferationoftheADSCs,which presentedsimilarresponseamongfivegroups.IncontrasttotheADSCspre-processedfor0day,theADSCspre-processed for3dayspresentedlowerexpressionoftypeIIcollagenandaggrecan;theADSCspre-processedfor7dayspresentedhigherexpressionoftypeIIcollagenandaggrecan;theADSCspre-processedfor10dayspresentedhigherexpressionoftypeIIcollagen,whilelowerexpressionofaggrecan;theADSCspre-processedfor14dayspresentedhigherexpressionoftypeIIcollagen,withoutsignificanceintheexpressionofaggrecan.TherewasnosignificantdifferenceamongthesefivegroupsintheexpressionoftypeIcollagen. Conclusion:TheresultsdemonstratethatADSCspre-processedinchondrogenesisdifferentialculturemediumforaboutoneweekmaybeabettercellsourceforthestemcelltherapyofthedegenerativediskdiseases.