hct-116、hela、MEFs细胞参数介绍.docVIP免费

HELAHeLa(ATCC®CCL-2™)ATCC：Hela细胞，源于黑人31岁女性，子宫颈腺癌，是一种附着型上皮细胞，要求存于液氮中。Thesecellsareasuitabletransfectionhost.ThiscelllinecanbeusedtoscreenforEscherichiacolistrainswithinvasivepotential.BiosafetyLevel：（2[Cellscontainhumanpapillomavirus]BiosafetyclassificationisbasedonU.S.PublicHealthServiceGuidelines,itistheresponsibilityofthecustomertoensurethattheirfacilitiescomplywithbiosafetyregulationsfortheirowncountry.）CompleteGrowthMedium：ThebasemediumforthiscelllineisATCC-formulatedEagle'sMinimumEssentialMedium,CatalogNo.30-2003.Tomakethecompletegrowthmedium,addthefollowingcomponentstothebasemedium:fetalbovineserumtoafinalconcentrationof10%.Subculturing（接种）：Volumesusedinthisprotocolarefora75cm2flask;proportionallyreduceorincreaseamountofdissociationmediumforculturevesselsofothersizes.CorningT-75flasks(catalog#430641)arerecommendedforsubculturingthisproduct.1.Removeanddiscardculturemedium.2.Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin-0.53mMEDTAsolutiontoremovealltracesofserumwhichcontainstrypsininhibitor.3.Add2.0to3.0mLofTrypsin-EDTAsolutiontoflaskandobservecellsunderaninvertedmicroscopeuntilcelllayerisdispersed(usuallywithin5to15minutes).Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthecellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.4.Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.5.Addappropriatealiquotsofthecellsuspensiontonewculturevessels.6.Incubateculturesat37C.SubcultivationRatio:Asubcultivationratioof1:2to1:6isrecommendedMediumRenewal:2to3timesperweekCryopreservationFreezeMedium:Completegrowthmediumsupplementedwith5%(v/v)DMSOStorageTemperature:LiquidnitrogenvaporphaseCultureConditionsAtmosphere:Air,95%;carbondioxide(CO2),5%Temperature:37°C论坛：贴壁生长，铺路石状，长得快，2-3天传代一次，传代不及时会造成老化的细胞堆积，看起来很脏。10%+1640或者10%+高糖DMEM都有培养。MEFs_283TAg(ATCC®CRL-2822™)OrganismMusmusculus,mouseTissueembryoCellTypefibroblastimmortalizedwithSV40largeTantigenSV40largeTantigentransfectedProductFormatfrozenMorphologyfibroblastCulturePropertiesadherentBiosafetyLevel2cellscontainingSV40viralDNAsequencesBiosafetyclassificationisbasedon聽U.S.PublicHealthServiceGuidelines,itistheresponsibilityofthecustomertoensurethattheirfacilitiescomplywithbiosafetyregulationsfortheirowncountry.Age14.5daygestationembryoApplicationsDNArepairstudiesStorageConditionsliquidnitrogenvaporphaseDerivation283TAg(Polb/Mpgdoublenull)isamouseembryonicfibroblast(MEF)celllinederivedfrompolymerase(DNAdirected),beta(Polb)/N-methylpurine-DNAglycosylase(Mpg,Aag)doublenullembryosatday14.5ofgestation.ThecelllinewasestablishedbytransfectionwithanexpressionvectorforSV40largeTantigen[PubMed:8538772].ThecellsaretransgenicforlambdaLIZ(LacI/cII).Comments283TAg(Polb/Mpgdoublenull)isamouseembryonicfibroblast(MEF)celllinederivedfrompolymerase(DNAdirected),beta(Polb)/N-methylpurine-DNAglycosylase(Mpg,Aag)doublenullembryosatday14.5ofgestation.ThecelllinewasestablishedbytransfectionwithanexpressionvectorforSV40largeTantigen[PubMed:8538772].ThecellsaretransgenicforlambdaLIZ(LacI/cII).CompleteGrowthMediumThebasemediumforthiscelllineisATCC-formulatedDulbecco'sModifiedEagle'sMedium,CatalogNo.30-2002.Tomakethecompletegrowthmedium,addthefollowingcomponentstothebasemedium:fet...