Description

ProductSheet

HCC827(ATCCCRL2868)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
1

Organism:Homosapiens,human
Tissue:lung
Disease:adenocarcinoma
CellType:epithelial
Age:39yearsadult
Gender:female
GrowthProperties:adherent
DNAProfile:
Amelogenin:X
CSF1PO:11
D13S317:9
D16S539:12
D5S818:12
D7S820:11,12
THO1:6
TPOX:8
vWA:18

BatchSpecificInformation
IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
RPMI1640Medium,ATCC302001.Tomakethe
completegrowthmedium,addthefollowing
componentstothebasemedium:fetalbovineserum
(ATCC302020)toafinalconcentrationof10%.

RefertotheCertificateofAnalysisforbatchspecifictestresults.

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

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

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HCC827(ATCCCRL2868)

HandlingProcedureforFrozenCells
Toinsurethehighestlevelofviability,thawthevialandinitiatethecultureassoonaspossibleuponreceipt.If
uponarrival,continuedstorageofthefrozencultureisnecessary,itshouldbestoredinliquidnitrogenvapor
phaseandnotat70C.Storageat70Cwillresultinlossofviability.
1. Thawthevialbygentleagitationina37Cwaterbath.Toreducethepossibilityofcontamination,keep
theOringandcapoutofthewater.Thawingshouldberapid(approximately2minutes).
2. Removethevialfromthewaterbathassoonasthecontentsarethawed,anddecontaminateby
dippinginorsprayingwith70%ethanol.Alloftheoperationsfromthispointonshouldbecarriedout
understrictasepticconditions.
3. Transferthevialcontentstoacentrifugetubecontaining9.0mLcompleteculturemediumandspinat
approximately125xgfor5to10minutes.
4. Resuspendcellpelletwiththerecommendedcompletemedium(seethespecificbatchinformationfor

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

Orcontactyourlocaldistributor

theculturerecommendeddilutionratio)anddispenseintoa25cm2ora75cm2cultureflask.Itis
importanttoavoidexcessivealkalinityofthemediumduringrecoveryofthecells.Itissuggestedthat,
priortotheadditionofthevialcontents,theculturevesselcontainingthecompletegrowthmediumbe
placedintotheincubatorforatleast15minutestoallowthemediumtoreachitsnormalpH(7.0to7.6).
5. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif
usingthemediumdescribedonthisproduct.

HandlingProcedureforFlaskCultures
Theflaskwasseededwithcells(seespecificbatchinformation),grown,andcompletelyfilledwithmediumat
ATCCtopreventlossofcellsduringshipping.
1. Uponreceipt,visuallyexaminethecultureformacroscopicevidenceofanymicrobialcontamination.

Page1of3

ProductSheet

HCC827(ATCCCRL2868)

Usinganinvertedmicroscope(preferablyequippedwithphasecontrastoptics),carefullycheckfor
anyevidenceofmicrobialcontamination.Also,checktodetermineifthemajorityofcellsarestill
attachedtothebottomoftheflaskduringshippingtheculturesaresometimeshandledroughlyand
manyofthecellsoftendetachandbecomesuspendedintheculturemedium(butarestillviable).
2. Ifthecellsarestillattached,asepticallyremoveallbut5to10mLoftheshippingmedium.The
shippingmediumcanbesavedforreuse.Incubatethecellsat37Cina5%CO2inairatmosphere
untiltheyarereadytobesubcultured.
3. Ifthecellsarenotattached,asepticallyremovetheentirecontentsoftheflaskandcentrifugeat
125xgfor5to10minutes.Removeshippingmediumandsave.Resuspendthepelletedcellsin10
mLofthismediumandaddto25cm2flask.Incubateat37Cina5%CO2inairatmosphereuntilcells

PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
1

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

arereadytobesubcultured.

SubculturingProcedure
Volumesusedinthisprotocolarefor75cm2flaskproportionallyreduceorincreaseamountofdissociation
mediumforculturevesselsofothersizes.
1. Removeanddiscardculturemedium.
2. Brieflyrinsethecelllayerwithor0.25%(w/v)Trypsin0.53mMEDTAsolutionorDulbeccos
PhosphateBufferedSaline(DPBS)toremovealltracesofserumthatcontainstrypsininhibitor.
3. Add1.0to3.0mLofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
untilthecelllayerisdispersed(usuallywithin5to15minutes).
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthe
cellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.
4. Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.
5. Transfercellsuspensiontocentrifugetubeandspinatapproximately125xgfor5to10minutes.
6. Discardsupernatantandresuspendcellsinfreshgrowthmedium.Addappropriatealiquotsofcell
suspensiontonewculturevessels.Aninoculumof5x103to7x103viablecells/cm2is
recommended.
7. Placeculturevesselsinincubatorsat37C.Maintainculturesatacellconcentrationbetween3x104

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
RPMI1640Medium,ATCC302001.Tomakethe
completegrowthmedium,addthefollowing
componentstothebasemedium:fetalbovineserum
(ATCC302020)toafinalconcentrationof10%.

and5x104cells/cm2.
SubcultivationRatio:Asubcultivationratioof1:4to1:6isrecommended
MediumRenewal:Every2to3days

CryopreservationMedium
Completegrowthmediumdescribedabovesupplementedwith5%(v/v)DMSO.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HCC827(ATCCCRL2868)

CellculturetestedDMSOisavailableasATCCCatalogNo.4X.

Comments
ThislungadenocarcinomahasanacquiredmutationintheEGFRtyrosinekinasedomain(E746A750
deletion).

References
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:1
Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

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

Orcontactyourlocaldistributor

Page2of3

ATCCproductsarewarrantedfor30daysfromthedateofshipment,andthiswarrantyisvalidonlyifthe
productisstoredandhandledaccordingtotheinformationincludedonthisproductinformationsheet.Ifthe
ATCCproductisalivingcellormicroorganism,ATCCliststhemediaformulationthathasbeenfoundtobe
effectiveforthisproduct.Whileother,unspecifiedmediamayalsoproducesatisfactoryresults,achangein
mediaortheabsenceofanadditivefromtheATCCrecommendedmediamayaffectrecovery,growthand/or
functionofthisproduct.Ifanalternativemediumformulationisused,theATCCwarrantyforviabilityisno
longervalid.

Disclaimers

ProductSheet

HCC827(ATCCCRL2868)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
1

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
RPMI1640Medium,ATCC302001.Tomakethe
completegrowthmedium,addthefollowing
componentstothebasemedium:fetalbovineserum
(ATCC302020)toafinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HCC827(ATCCCRL2868)

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

Orcontactyourlocaldistributor

Page3of3

Thisproductisintendedforlaboratoryresearchpurposesonly.Itisnotintendedforuseinhumans.
WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproductsheet,
ATCCmakesnowarrantiesorrepresentationsastoitsaccuracy.Citationsfromscientificliteratureand
patentsareprovidedforinformationalpurposesonly.ATCCdoesnotwarrantthatsuchinformationhasbeen
confirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,anduse.ATCC
isnotliableforanydamagesorinjuriesarisingfromreceiptand/oruseofthisproduct.Whilereasonableeffort
ismadetoinsureauthenticityandreliabilityofmaterialsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofsuchmaterials.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2014.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[10/30]