Description

ProductSheet

MCF7(ATCCHTB22)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
1

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
Eagle'sMinimumEssentialMedium,CatalogNo.30
2003.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:0.01mg/ml
humanrecombinantinsulinfetalbovineserumtoa
finalconcentrationof10%.

Organism:Homosapiens,human
Tissue:mammarygland,breastderivedfrommetastaticsite:pleuraleffusion
Disease:adenocarcinoma
CellType:epithelial
Age:69yearsadult
Gender:female
Morphology:epithelial
GrowthProperties:adherent
Isoenzymes:
AK1,1
ESD,12
G6PD,B
GLOI,12
PGM1,12
PGM3,1
DNAProfile:
Amelogenin:X
CSF1PO:10
D13S317:11
D16S539:11,12
D5S818:11,12
D7S820:8,9
THO1:6
TPOX:9,12
vWA:14,15
CytogeneticAnalysis:modalnumber=82range=66to87.
Thestemlinechromosomenumbersrangedfromhypertriploidytohypotetraploidy,withthe2Scomponent
occurringat1%.Therewere29to34markerchromosomesperSmetaphase24to28markersoccurredinat
least30%ofcells,andgenerallyonelargesubmetacentric(M1)and3largesubtelocentric(M2,M3,andM4)
markerswererecognizableinover80%ofmetaphases.NoDMweredetected.Chromosome20was
nullisomicandXwasdisomic.

BatchSpecificInformation
RefertotheCertificateofAnalysisforbatchspecifictestresults.

CitationofStrain

SAFETYPRECAUTION
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:MCF7(ATCCHTB22)

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. Transferthecellpellettoanappropriatesizevessel.Itisimportanttoavoidexcessivealkalinityofthe
mediumduringrecoveryofthecells.Itissuggestedthat,priortotheadditionofthevialcontents,the
culturevesselcontainingthegrowthmediumbeplacedintotheincubatorforatleast15minutesto
allowthemediumtoreachitsnormalpH(7.0to7.6).
5. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif

ProductSheet

MCF7(ATCCHTB22)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
1

usingthemediumdescribedonthisproductsheet.

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.

IntendedUse

SubculturingProcedure
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
Eagle'sMinimumEssentialMedium,CatalogNo.30
2003.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:0.01mg/ml
humanrecombinantinsulinfetalbovineserumtoa
finalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:MCF7(ATCCHTB22)

Volumesusedinthisprotocolarefor75cm2flasksproportionallyreduceorincreaseamountofdissociation
mediumforculturevesselsofothersizes.CorningT75flasks(catalog#430641)arerecommendedfor
subculturingthisproduct.
Note:iffloatingcellsarepresent,itisrecommendedthattheybetransferredatthefirsttwo(2)subcultures
asdescribedbelow.Itisnotnecessarytotransferfloatingcellsforsubsequentsubcultures.
1. Removeculturemediumtoacentrifugetube.
2. Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin0.53mMEDTAsolutiontoremovealltracesof
serumwhichcontainstrypsininhibitor.
3. Add2.0to3.0mLofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
untilcelllayerisdispersed(usuallywithin5to15minutes).
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthe
cellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.
4. Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.
5. Transferthecellsuspensiontothecentrifugetubewiththemediumandcellsfromstep1,and
centrifugeatapproximately125xgfor5to10minutes.Discardthesupernatant.
6. Resuspendthecellpelletinfreshgrowthmedium.Addappropriatealiquotsofthecellsuspensionto
newculturevessels.
7. Incubateculturesat37C.
SubcultivationRatio:Asubcultivationratioof1:3to1:6isrecommended
MediumRenewal:2to3timesperweek

CryopreservationMedium
Completegrowthmediumdescribedabovesupplementedwith5%(v/v)DMSO.CellculturetestedDMSOis
availableasATCCCatalogNo.4X.

Comments

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

Orcontactyourlocaldistributor

Page2of3

TheMCF7lineretainsseveralcharacteristicsofdifferentiatedmammaryepitheliumincludingabilitytoprocess
estradiolviacytoplasmicestrogenreceptorsandthecapabilityofformingdomes.Thecellsexpressthe
WNT7Boncogene.
ref
GrowthofMCF7cellsisinhibitedbytumornecrosisfactoralpha(TNFalpha).
SecretionofIGFBP'scanbemodulatedbytreatmentwithantiestrogens.

References
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:1

ProductSheet

MCF7(ATCCHTB22)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

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

Disclaimers

BiosafetyLevel
1

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
Eagle'sMinimumEssentialMedium,CatalogNo.30
2003.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:0.01mg/ml
humanrecombinantinsulinfetalbovineserumtoa
finalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:MCF7(ATCCHTB22)

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
ismadetoensureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2014.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[08/20]