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《差速貼壁技術(shù)對(duì)大鼠腦皮質(zhì)星形膠質(zhì)細(xì)胞純化率的影響》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、周欣,明曉云,康頌建,白波,段耀奎.差速貼壁技術(shù)對(duì)大鼠腦皮質(zhì)星形膠質(zhì)細(xì)胞純化率的影響.中國(guó)組織工程研究與臨床康復(fù),2007,11(15):2829-2831.InfluenceofdifferentialvelocityadherenttechniqueonthepurifiedrateofcerebralastrocytesinratsAbstractAIM:Toobservetheproportionofratcerebralastrocyteculturedbydifferentialvelocityadherenttechnique,soas
2、toestablishasetofreliabletechniquefortheisolationandpurificationoftheastrocytescerebralcortex.METHODS:TheexperimentwasperformedintheInstituteofLifeScience,TaishanMedicalCollegefromJunetoAugust2006.Wistarratsaged2-3daysafterbirthofeithersexwereprovidedbyExperimentalAnimalCenter,
3、InstituteofLifeScience,TaishanMedicalCollege,andselectedtocarryoutprimarycultureofcerebralastrocytes.Theyweredividedintotwogroups:routineculturegroupanddifferentialvelocityadherentgroup.Astrocyteswereremovedatminutes15and30inthelattergroup,andthencultureflaskwasrotatedtoremoves
4、upernatantintoanothercultureflask,andthenkeptinincubatorforfollowingculture.After7-10days,eachgroupwaspassagedtillcellsgrewintodemixing,andthenputinswingbedat37℃andshookat250r/minfor18hours.Supernatantwasadded,andthenthecellswerewashedwithD-Hank'sliquorthreetimes,andtreatedwith
5、0.25%trypsinization,observedunderinvertedmicroscope.Mediumcontainingserumwasaddedtostopdigestionwhenprocessofcellsrecovered.Thecellswereblownandhitbypipetteforkeepingcellsfromflaskwall,andthencentrifugedat1000r/minfor5minutes.SupernatantwasremovedbeforeDMEMmediumcontaining0.2vo
6、lumefractionwassuspendedandprecipitated.ThecellswereincubatedinL-polylysinecultureflasksuccessively.Doubleimmunofluorescencestainingwasselectedtoidentifypurificationoftheastrocytes.Atthesametime,integratedopticaldensity(IOD)wasusedtomeasuregrowthstateofastrocytes.RESULTS:①Purit
7、yofastrocytescouldbeelevatedmarkedlybydifferentialvelocityadherenttechnique[routineculturegroup:(82±3)%,differentialvelocityadherentgroupof15minutes:(94±2)%,differentialvelocityadherentgroupof30minutes:(95±2)%,P<0.01.Differentialvelocityadherenttechniqueneededenoughtime.Therewa
8、snosignificantdifferenceinelevatingcellpurityinthe15mi
