Description

ProductSheet

HEp2(ATCCCCL23)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

Organism:Homosapiens,human
Tissue:HeLacontaminant
Disease:Carcinoma
Morphology:epithelial
HelaMarkers:Y
GrowthProperties:adherent
VirusSusceptibility:
ViralTesting:ATCCconfirmedthiscelllineispositiveforthepresenceofhumanpapillomaviralDNA
sequencesviaPCR.
Isoenzymes:
G6PD,A
DNAProfile:
Amelogenin:X
CSF1PO:9,10
D13S317:12,13.3
D16S539:9,10
D5S818:11,12
D7S820:8,12
THO1:7
TPOX:8,12
vWA:16,18
CytogeneticAnalysis:Occasionalpolyploids.Severalmarkerchromosomeswereobservedalongwith
frequentminutes,andoften2largechromosomeswithsubterminalcentromeres.HeLaMarkerChromosomes:
OnecopyofM2,twofourcopiesofM3andonecopyofM4asrevealedbyGbandingpatterns.

BatchSpecificInformation
CompleteGrowthMedium
RefertotheCertificateofAnalysisforbatchspecifictestresults.
ThebasemediumforthiscelllineisATCCformulated
Eagle'sMinimumEssentialMedium,CatalogNo.30
2003.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:fetalbovine
serumtoafinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HEp2(ATCCCCL23)

SAFETYPRECAUTION
ATCChighlyrecommendsthatprotectiveglovesandclothingalwaysbeusedandafullfacemaskalwaysbe
wornwhenhandlingfrozenvials.Itisimportanttonotethatsomevialsleakwhensubmersedinliquidnitrogen
andwillslowlyfillwithliquidnitrogen.Uponthawing,theconversionoftheliquidnitrogenbacktoitsgas
phasemayresultinthevesselexplodingorblowingoffitscapwithdangerousforcecreatingflyingdebris.

Unpacking&StorageInstructions
1. Checkallcontainersforleakageorbreakage.
2. Removethefrozencellsfromthedryicepackagingandimmediatelyplacethecellsatatemperature
below130C,preferablyinliquidnitrogenvapor,untilreadyforuse.

HandlingProcedureforFrozenCells

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

Page1of3

Toinsurethehighestlevelofviability,thawthevialandinitiatethecultureassoonaspossibleuponreceipt.If
uponarrival,continuedstorageofthefrozencultureisnecessary,itshouldbestoredinliquidnitrogenvapor
phaseandnotat70C.Storageat70Cwillresultinlossofviability.
1. Thawthevialbygentleagitationina37Cwaterbath.Toreducethepossibilityofcontamination,keep
theOringandcapoutofthewater.Thawingshouldberapid(approximately2minutes).
2. Removethevialfromthewaterbathassoonasthecontentsarethawed,anddecontaminateby
dippinginorsprayingwith70%ethanol.Alloftheoperationsfromthispointonshouldbecarriedout
understrictasepticconditions.
3. Itisrecommendedthatthecryoprotectiveagentberemovedimmediately.Centrifugethecell
suspensionatapproximately125xgfor5to10minutes.Discardthesupernatantandresuspendthe
cellpelletinanappropriateamountoffreshgrowthmedium.
4. Transferthevialcontentstoanappropriatesizevessel.Itisimportanttoavoidexcessivealkalinityof
themediumduringrecoveryofthecells.Itissuggestedthat,priortotheadditionofthevialcontents,
theculturevesselcontainingthegrowthmediumbeplacedintotheincubatorforatleast15minutesto
allowthemediumtoreachitsnormalpH(7.0to7.6).
5. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif
usingthemediumdescribedonthisproductsheet.

HandlingProcedureforFlaskCultures

ProductSheet

HEp2(ATCCCCL23)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

Theflaskwasseededwithcells(seespecificbatchinformation)grownandcompletelyfilledwithmediumat
ATCCtopreventlossofcellsduringshipping.
1. Uponreceiptvisuallyexaminethecultureformacroscopicevidenceofanymicrobialcontamination.
Usinganinvertedmicroscope(preferablyequippedwithphasecontrastoptics),carefullycheckfor
anyevidenceofmicrobialcontamination.Alsochecktodetermineifthemajorityofcellsarestill
attachedtothebottomoftheflaskduringshippingtheculturesaresometimeshandledroughlyand
manyofthecellsoftendetachandbecomesuspendedintheculturemedium(butarestillviable).
2. Ifthecellsarestillattached,asepticallyremoveallbut5to10mLoftheshippingmedium.The
shippingmediumcanbesavedforreuse.Incubatethecellsat37Cina5%CO2inairatmosphere
untiltheyarereadytobesubcultured.
3. Ifthecellsarenotattached,asepticallyremovetheentirecontentsoftheflaskandcentrifugeat
125xgfor5to10minutes.Removeshippingmediumandsave.Resuspendthepelletedcellsin10
mLofthismediumandaddto25cm2flask.Incubateat37Cina5%CO2inairatmosphereuntilcells
arereadytobesubcultured.

SubculturingProcedure

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
Eagle'sMinimumEssentialMedium,CatalogNo.30
2003.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:fetalbovine
serumtoafinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing

Volumesusedinthisprotocolarefor75cm2flaskproportionallyreduceorincreaseamountofdissociation
mediumforculturevesselsofothersizes.
1. Removeanddiscardculturemedium
2. Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin0.53mMEDTAsolutiontoremovealltracesof
serumwhichcontainstrypsininhibitor.
3. Add2.0to3.0mLofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
untilcelllayerisdispersed(usuallywithin5to15minutes).Note:Toavoidclumpingdonotagitatethe
cellsbyhittingorshakingtheflaskwhilewaitingforthecellstodetach.Cellsthataredifficultto
detachmaybeplacedat37Ctofacilitatedispersal.
4. Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.
5. Addappropriatealiquotsofthecellsuspensiontonewculturevessels.
6. Incubateculturesat37C.
SubcultivationRatio:Asubcultivationratioof1:4to1:10isrecommended
MediumRenewal:2to3timesperweek

CryopreservationMedium
Completeculturemediumdescribedabovesupplementedwith5%(v/v)DMSO.CellculturetestedDMSOis
availableasATCCCatalogNo.4X.

Comments

manner:HEp2(ATCCCCL23)

CellsofthislinecontainHeLamarkerchromosomes,andwerederivedviaHeLacontamination.Thislinewas
originallythoughttobederivedfromanepidermoidcarcinomaofthelarynx,butwassubsequentlyfound,
basedonisoenzymeanalysis,HeLamarkerchromosomes,andDNAfingerprinting,tohavebeenestablished
viaHeLacellcontamination.Thecellsarepositiveforkeratinbyimmunoperoxidasestaining.ATCCconfirmed
thiscelllineispositiveforthepresenceofhumanpapillomaviralDNAsequencesviaPCR.

References
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:2

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

Page2of3

Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty
TheviabilityofATCCproductsiswarrantedfor30daysfromthedateofshipment,andisvalidonlyifthe
productisstoredandculturedaccordingtotheinformationincludedonthisproductinformationsheet.ATCC
liststhemediaformulationthathasbeenfoundtobeeffectiveforthisstrain.Whileother,unspecifiedmedia
mayalsoproducesatisfactoryresults,achangeinmediaortheabsenceofanadditivefromtheATCC

recommendedmediamayaffectrecovery,growthand/orfunctionofthisstrain.Ifanalternativemedium
formulationisused,theATCCwarrantyforviabilityisnolongervalid.

Disclaimers

ProductSheet

HEp2(ATCCCCL23)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

Thisproductisintendedforlaboratoryresearchpurposesonly.Itisnotintendedforuseinhumans.
WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproductsheet,
ATCCmakesnowarrantiesorrepresentationsastoitsaccuracy.Citationsfromscientificliteratureand
patentsareprovidedforinformationalpurposesonly.ATCCdoesnotwarrantthatsuchinformationhasbeen
confirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,anduse.ATCC
isnotliableforanydamagesorinjuriesarisingfromreceiptand/oruseofthisproduct.Whilereasonableeffort
ismadetoinsureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2013.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[07/19]

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
Eagle'sMinimumEssentialMedium,CatalogNo.30
2003.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:fetalbovine
serumtoafinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HEp2(ATCCCCL23)

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

Page3of3