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HELAHeLa ATCC©CCL-2细胞，源于黑人岁女性，子宫颈腺癌，是一种附着型上皮细胞，要求存ATCC Hela31于液氮中These cells are a suitable transfection host.This cell line canbe usedto screenforEscherichia colistrains withinvasive potential.Biosafety Level2[Cells containhuman papillomavirus]Biosafetyclassification is based on U.S.Public HealthService Guidelines,it isthe responsibilityof thecustomer to ensure that theirfacilities complywith biosafetyregulations fbrtheir owncountry.Complete GrowthMedium Thebase medium for thiscell line is ATCC-fbrmulated EaglesMinimumEssential Medium,Catalog No.30-
2003.To makethe complete growth medium,addthe following components to the base medium:fetal bovineserum to a final concentration of10%.Subculturing接种Volumes usedin thisprotocol arefor a75cm2flask;proportionally reduceor increaseamountof dissociationmediumforculture vesselsof othersizes.CorningT-75flasks catalog#430641arerecommended for subculturing this product.
1.Remove anddiscard culturemedium.
2.Briefly rinsethe celllayer with
0.25%w/v Trypsin-
0.53mM EDTA solution to remove all traces of serumwhich containstrypsin inhibitor.
3.Add
2.0to
3.0mL ofTrypsin-EDTA solutionto flaskand observecells under an inverted microscope until celllayer isdispersed usuallywithin5to15minutes.Note:To avoidclumping donot agitatethe cellsby hittingor shakingthe flask while waiting for the cells todetach.Cells that are difficult to detach may be placed at37C to facilitate dispersal.
4.Add
6.0to
8.0mL ofcomplete growth medium andaspirate cellsby gently pipetting.
5.Add appropriatealiquots of the cellsuspension tonew culture vessels.
6.Incubate culturesat37C.Subcultivation Ratio:A subcultivationratio of1:2to1:6is recommendedMediumRenewal:2to3times perweekCryopreservation FreezeMedium:Complete growthmedium supplementedwith5%v/v DMSOStorageTemperature:Liquid nitrogen vapor phaseCultureConditionsAir,95%;carbon dioxideC02,5%Atmosphere:37°CTemperature:论坛贴壁生长，铺路石状，长得快，天传代一次，传代不及时会造成老化的细胞堆2-3积，看起来很脏或者高糖都有培养10%+164010%+DMEMMEFs_283TAg ATCC®CRL-2822™Organism Musmusculus,mouseembryo fibroblastimmortalized withSV40large TantigenSV40large Tantigen transfectedTissuefrozenfibroblastCell TypeCultureProperties adherentProductFormatMorphologyBiosafety Level2cells containingSV40viral DNAsequences醵Biosafety classification isbased onU.S.Public HealthServiceGuidelines,it isthe responsibilityof thecustomer toensure thattheirfacilities complywith biosafetyregulations fortheir owncountry.Age
14.5day gestationembryoApplications DNArepair studiesStorage Conditions liquid nitrogenvaporphaseDerivation283TAg Polb/Mpg doublenull isa mouseembryonic fibroblastMEFcell linederived frompolymerase DNAdirected,beta Polb/N-methylpurine-DNA glycosylaseMpg,Aag doublenull embryosatday
14.5of gestation.The cell line wasestablished bytransfectionwith anexpression vectorfor SV40large Tantigen[PubMed:8538772].The cellsare transgenicfor lambdaLIZ Lacl/cll.Comments283TAg Polb/Mpg doublenull isa mouseembryonic fibroblastMEFcell linederived frompolymerase DNAdirected,beta Polb/N-methylpurine-DNA glycosylaseMpg,Aag doublenull embryosatday
14.5of gestation.The cellline wasestablished bytransfectionwith anexpression vectorfor SV40large Tantigen[PubMed:8538772].The cellsare transgenicfor lambdaLIZ Lacl/cll.Complete GrowthMedium Thebase mediumfor thiscelllineis ATCC-formulated DulbeccosModifiedEagles Medium,Catalog No.30-
2002.To makethe completegrowthmedium,add thefollowingcomponents tothe base medium:fetalbovine serum toa finalconcentration of10%.Subculturing Protocol:
1.Remove anddiscard culturemedium.
2.Briefly rinsethe celllayer with
0.25%w/v Trypsin-
0.53mM EDTAsolutionto removeall tracesof serumthat containstrypsin inhibitor.
3.Add
2.0to
3.0ml ofTrypsin-EDTA solutionto flaskand observecellsunderaninverted microscopeuntil celllayer isdispersedusually within5to15minutes.Note:To avoidclumping donot agitatethe cellsby hittingorshaking theflaskwhilewaiting forthe cellsto detach.Cells that are℃difficultto detachmay beplaced at37tofacilitate dispersal.
4.Add
6.0to
8.0ml ofcomplete growthmedium andaspiratecells bygentlypipetting.
5.Add appropriatealiquots of the cellsuspension tonew culturevessels.An inoculumof4X103to4X104viable cells/cm2isrecommended.℃
6.Incubate culturesat
37.Interval:Maintain culturesatacell concentrationbetween6X103and1X105cells/cm
2.Subcultivation Ratio:A subcultivationratio of1:6to1:8isrecommendedMedium Renewal:Two tothree timesweeklyCryopreservation Freeze medium:Complete growthmedium supplementedwith5%v/vDMSOStorage temperature:liquid nitrogenvapor phaseCultureConditions Atmosphere:air,95%;carbon dioxideCO2,5%℃Temperature:
37.0Population DoublingTime29hoursHCT116ATCC@CCL-247™Organism Homosapiens,humanTissue colonProductFormat frozenMorphologyepithelialCulture PropertiesadherentBiosafety LevelBiosafetyclassificationisbasedonU.S.Public HealthServiceGuidelines,^it isthe responsibilityof thecustomertoensurethattheirfacilities complywith biosafetyregulations fortheir owncountry.结肠直肠癌Disease colorectalcarcinomaAge adultGendermaleApplicationsThis celllineisasuitabletransfectionhost.This linehas amutation incodon13of theras proto-oncogene,and canbeused asa positivecontrol forPCR assaysof mutationin thiscodon.StorageConditionsliquid nitrogenvapor phaseKaryotypeThe stemlinechromosome numberis neardiploid withthe modalnumberat4562%and polyploidsoccurring at
6.8%.The markers10q+and t8p;18q arepresent inall metaphasesand t9q;16p-,in80%ofthe cells karyotyped.N16is monosomicin thepresence of,butdisomic inthe absenceoft9q;16p-.N10and N18are monosomicandother chromosomesfrom thosementioned aboveare disomic.Q-band observationsrevealed thepresence ofthe Ychromosome,butnot inall cells50%of cellslacked theY inG-band karyotypes.ImagesClinical DatamaleGenes Expressedcarcinoembryonic antigenCEA1ng per106cells per10days.Cellular Productscarcinoembryonic antigenCEA1ng per10exp6cells per10days;keratinTumorigenic YesYes,in nudemiceEffectsRefCommentsThe cellsare positive for keratinby immunoperoxidasestaining.HCT116cellsarepositivefortransforming growthfactor beta1TGFbeta1and beta2TGF beta2expression.Complete GrowthMedium Thebasemediumfor thiscelllineis ATCC-formulated McCoys5a MediumModified,Catalog No.30-
2007.To makethe completegrowthmedium,add thefollowingcomponentstothebasemedium:fetal bovineserumtoafinal辗concentrationof10%Subculturing Volumesare givenfor a75cm2B flask.Corning庐H T-75flasks catalog#430641are recommendedforsubculturingthisproduct.Increase ordecreasethe amountof dissociationmedium neededproportionally forculturevesselsofother sizes.
1.Remove anddiscard culturemedium.
2.Briefly rinsethe celllayer with
0.25%w/v Trypsin-
0.53mM EDTAsolutiontoremovealltracesofserumwhich containstrypsin inhibitor.
3.Add
2.0to
3.0mL ofTrypsin-EDTAsolutionto flaskand observecellsunder aninvertedmicroscopeuntilcelllayer isdispersed usuallywithin5to15minutes.Note:To avoidclumping donot agitatethe cellsby hittingor shakingthe flaskwhilewaitingforthecellsto detach.Cells thatare difficulttodetachmaybe掳placedat37C tofacilitatedispersal.
4.Add
6.0to
8.0mL ofcompletegrowthmedium andaspirate cellsbygently pipetting.
5.Add appropriatealiquots ofthecellsuspension tonew culturevessels.掳
6.Incubate culturesat37C.疆Subcultivation Ratio:A subcultivationratio of1:3to1:8is recommendedMediumRenewal:^Every2to3days糜FreezemediumComplete growthmedium supplementedwith5%v/vDMSOStorage temperature:^liquidnitrogenvapor phaseCryopreservationAtmosphere:^air,95%;carbon dioxideCO2,5%掳Temperature:^37CGrowth Conditions:^!Growth andplating efficiencyare enhancedby usingaCulture Conditionsfeederlayer ofmurine fibroblasts.人结肠癌细胞是等人在年从一例男性结肠癌患者的癌组H CT—116B ra ttai n1979织中培养建立起来的低分化型腺癌细胞株［］,广泛应用于恶性肿瘤细胞的生物学特性、1抗肿瘤药物作用机理和抗癌药物的筛选等领域的研究近年的研究表明，大多数HCT—1细胞具有肿瘤干细胞的特性，可作为肿瘤干细胞研究的理想对象［］在体外培养条166件下，细胞一般呈梭形或多角形贴壁生长，传代培养时通常采用胰蛋白酶HCT—116消化法进行内容总结1HELAHeLa ATCC@CCL-2™细胞，源于黑人岁女性，子宫颈腺癌，是一种附着型上ATCC Hela31皮细胞，要求存于液氮中。
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