体外三维支架诱导脂肪干细胞向髓核样细胞的分化

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1、万方数据中国组织工程研究与临床康复第13卷笫38期2009—09—17出版JournalofClinicalRehabilitativeTissueEngmeeringResearchV01．13,No．38疋尼露麴蝴磁体外三维支架诱导脂肪干细胞向髓核样细胞的分化术★Differentiationofadipose-derivedstemcellstowardsnucleuspulposus-likecellsinducedbythree-dimensionalscaffoldinvitroTianHai-quan，LiFangAbstractBACKGROUND：Thenucleuspu

2、lpoSUStissueengineeringisregardedasanideelmethodinrepairingintervertebraIdiscdegeneration．However，theresourceofnucleuspulposuscellsislimited，soseedcellplaysavitalroleinnucleuspulposus(NP)tissueengineering．OBJECTlVE：Toverifywhetheradipose-derivedstemcells(ADSCs)candifferentiateintoNP．1ikecellsinduc

3、edbythree-dimensionaIscaffold．DESIGN．TIMEANDSEl_rING：An／nv／troobservationbasedoncell．materialsIevelwasperformedattheOrthopaedicResearchInstitute．GeneraIHospitalofBeijingMilitaryAreaCommandofChinesePLAfromJulytoDecember2008．MATERIALS：ThteeJapanesebig．earedrabbitswith3-monthsoldwereprovidedbvVitaIRi

4、verLaboratoryAnimaITechnologyCo．，Ltd．，andthemedicalchitosanwaspurchasedfromGolden-shellBiochemicaICo．。Ltd．METHODS：TheADSCswereisolatedfrOmtherabbitadiposeandthensubculturedwithcollagenasedigestion．ThesecondADSCswereprepared50ULcellsuspension(5x106cells)，andmixedwith150ULchitosan—alginate(C／A)andcu

5、lturedina12-wellplatefor20minutesinroomtemperaturetoobtainG俄gel．Theexperimentdividedinto3groups，inthecontrolgmup。2mLDMEMcontaining1O％fetalbovineserumwasadded；DMEMcontainingbovineserumalbumin，transforminggro、Ivthfactor-1(TGF)，hexadecadrol，vitaminC，bonemorphogeneticprotein-7，1％lTSwasadded，andthencul

6、turedwith2％02or20％O2．A¨thesampleswereplacedinincubatoratahumidifiedatmosphereof5％02at37℃for21days．MAINOUTCOMEMEASURES：Histomorphologicalobservation；levelsofproteoglycan．aswellastypeIIcollagen．RESULTS：ThescanningelectronmicroscopyshowedthatADSCsgrewwelIonthechitosan．alginatescaffolds．whichwaspositi

7、vetooil-OstainingandAlcianbluestaining．Moreproteoglycanwasproduced．Withtimeincreasing，therewasnosignificantchangeintheproteoglycancontentandtypeIIcollageninthecontrolgroup．Thecontentofproteoglycanandcollagentype1