Adultbonemarrowstromalcellsculturedinvitroandinducedtodifferentiateintoosteoblasts:TengYong,HUYun,AnShujie,DaixianWen,ZhaoLiBaiJianping,LIXushengLugeonKeywords:]stemcells[Abstract]AIM:Tobuildamethodtoculturebonemarrowderivedmesenchymalstemcells(MSCs)invitroandtoinvestigatethefeasibilityofinducingMSCsintoosteoblasts.METHODS:HumanofMSCswereisolatedfromadultbonemarrowandpurifiedbybythepercollwereexpandedfordensitygradientcentrifugationandculturedinvitro.TheMSCsattachmentformedafter7-10dandtheMSCsofthepassage3werechosentoinduceintoosteoblasts.Fifteendayslater,thealkalinephosphataseassaywasconductedusingmodifiedcalciumcobaltstainingmethod,typeⅠcollagenandosteocalcinassaywasconductedusingimmunohistochemistryandcalciumnodeassaywasdoneusingalizarinredstaining.RESULTS:HomogeneousMSCswereobtainedbypercolldensitygradientcentrifugation.TheinducedMSCshadtypicalappearanceofosteoblasts.1TherateofALPexpressionwas81%,theexpressionsofcollagentypeⅠandosteocalcinwerepositive,andcalciumnodeswereseen.CONCLUSION:TheMSCscanbeseparatedfromadultbonemarrowandculturedinvitro,whichcanbeinducedintoosteoblasts.[Keywords]bonemarrow;stemcells;adult;tissueengineering;osteogenesis/drugeffects[Abstract]Objective:Toestablishadultbonemarrowstromalstemcells(MSCs)invitro,andexploretheorientationinduceddifferentiationasosteoblastsofways.Methods:extractadultbonemarrowPercolldensitygradientcentrifugationmethodvitroculturestickerswallcellpassaging3rdgenerationofcellsintheculturemediumintoboneinducerdexamethasone,βglycerophosphate,vitaminCandculturedfor15dintheinvertedmicroscopecellmorphology,calcium-cobaltstainingdetectionofalkalinephosphatase(ALPexpressionimmunohistochemistry(SABCassayofcollagentypeI,osteocalcinexpression,AlizarinRedstainingcalciumnoduleformation.:PercolldensitygradientcentrifugationcultureyieldedhomogeneousMSCsinduced2differentiationofMSCswastypicalosteogeniccellmorphology,theALPstainingpositiverateofupto81%ormore,positiveexpressionofcollagentypeI,osteocalcin,AlizarinRedstainingcalciumnoduleformationConclusion:youcanbeculturedfromadultbonemarrowMSCs,andcanbeinducedintoosteoblasts,cellscanbeusedasaclinicalbonetissueengineeringseed.[Keywords]bonemarrowstemcells;adults,tissueengineering,boneformation/drugeffects0IntroductionSeedcells,scaffolds,growthfactorsarethethreeelementsoftissueengineering,tofindasuitableseedcells[1]bonetissueengineeringseedcellsforbone,periosteum,bonemarrowandothernon-bonetissuemarrowisanimportantfactorforsuccessstromalstemcells(marrowmesenchymalstemcellsofMSCsitsadequatesources,drawnconvenient,safe,easytotrainandgeneticallymodified,hyperplasia,multi-differentiationandosteogenictheexactcapacity[2-4],whichattractedwidespreadattention,withawiderange3ofapplicationsprospects.SinceManiatopoulosetal[5]in1988firstreportedratMSCsculturedinvitrosincetheformationofcalcifiedbone-liketissue,bonemarrowfromanimalinducedtodifferentiateintobonecellsisnotdifficult,ithasbeenisolatedfromfetalbonemarrowMSCs.adultbonemarrowMSCscontentlesswhetherseparationseparationmethodishowthedirectionofthecurrentstudytheclinicaladultMSCsculturedandinducedosteogenicthisstudyonthebasisofanimalexperiments,topreparefortheorganizationworkstowardclinical.1Materialsandmethods1.1MaterialsBonemarrowtakenfromthefivecasesoforthopedicpatientswithiliac,aged28,30,35,55,60yearsofage.2patientswerediagnosedwithspondylolist...