Description

Organism:Musmusculus,mouse Strain:BALB/cfC3H Tissue:mammarygland Disease:ThistumorisananimalstageIVhumanbreastcancer. Morphology:epithelial GrowthProperties:adherent

ProductSheet

4T1(ATCCCRL2539)
PleasereadthisFIRST

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

StorageTemp. liquidnitrogen vapor temperature

Unpacking&StorageInstructions

BiosafetyLevel 1

1. Checkallcontainersforleakageorbreakage. 2. Removethefrozencellsfromthedryicepackagingandimmediatelyplacethecellsatatemperature below130C,preferablyinliquidnitrogenvapor,untilreadyforuse.

HandlingProcedureforFrozenCells
IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot intendedforanyanimalorhumantherapeuticor diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated RPMI1640Medium,CatalogNo.302001.Tomakethe completegrowthmedium,addthefollowing componentstothebasemedium:fetalbovineserumto afinalconcentrationof10%.

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

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it shouldbecitedinthatmanuscriptinthefollowing manner:4T1(ATCCCRL2539)

HandlingProcedureforFlaskCultures
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.

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

SubculturingProcedure
NOTE:thecellsshouldnotbeallowedtobecomeconfluent,subcultureat80%ofconfluence.Remove medium,andrinsewith0.25%trypsin0.53mMEDTAsolution.Removethesolutionandaddanadditional1to2 mLoftrypsinEDTAsolution.Allowtheflasktositatroomtemperature(orat37C)untilthecellsdetach.Add

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freshculturemedium,aspirateanddispenseintonewcultureflasks. SubcultivationRatio:Asubcultivationratioof1:6to1:8isrecommended MediumRenewal:Every2to3days

CryopreservationMedium

ProductSheet

4T1(ATCCCRL2539)
PleasereadthisFIRST

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

Comments
WheninjectedintoBALB/cmice,4T1spontaneouslyproduceshighlymetastatictumorsthatcanmetastasize tothelung,liver,lymphnodesandbrainwhiletheprimarytumorisgrowinginsitu. Theprimarytumordoesnothavetoberemovedtoinducemetastaticgrowth. Because4T1isresistantto6thioquanine,micrometastaticcells(asfewas1)canbedetectedinmany distantsiteorganswithbetteraccuracythatmosttumormodels.Thereisnoneedtocountnodulesorweight targetorgans.

StorageTemp. liquidnitrogen vapor temperature

References

BiosafetyLevel 1

Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:1
IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot intendedforanyanimalorhumantherapeuticor diagnosticuse.

Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S. DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes forHealth.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated RPMI1640Medium,CatalogNo.302001.Tomakethe completegrowthmedium,addthefollowing componentstothebasemedium:fetalbovineserumto afinalconcentrationof10%.

ATCCWarranty
TheviabilityofATCCproductsiswarrantedfor30daysfromthedateofshipment,andisvalidonlyifthe productisstoredandculturedaccordingtotheinformationincludedonthisproductinformationsheet.ATCC liststhemediaformulationthathasbeenfoundtobeeffectiveforthisstrain.Whileother,unspecifiedmedia mayalsoproducesatisfactoryresults,achangeinmediaortheabsenceofanadditivefromtheATCC recommendedmediamayaffectrecovery,growthand/orfunctionofthisstrain.Ifanalternativemedium formulationisused,theATCCwarrantyforviabilityisnolongervalid.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it shouldbecitedinthatmanuscriptinthefollowing manner:4T1(ATCCCRL2539)

Disclaimers
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.[05/31]

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

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