Description

Organism:Musmusculus,mouse
Tissue:
ProductSheet embryo
CellType:fibroblast
3T3L1(ATCCCL173) Age:embryo
Morphology:fibroblast
GrowthProperties:adherent
PleasereadthisFIRST
BatchSpecificInformation
StorageTemp.
RefertotheCertificateofAnalysisforbatchspecifictestresults.
liquidnitrogen
vaporphase
SAFETYPRECAUTION

ATCChighlyrecommendsthatprotectiveglovesandclothingalwaysbeusedandafullfacemaskalwaysbe
BiosafetyLevel wornwhenhandlingfrozenvials.Itisimportanttonotethatsomevialsleakwhensubmersedinliquidnitrogen

1 andwillslowlyfillwithliquidnitrogen.Uponthawing,theconversionoftheliquidnitrogenbacktoitsgas
phasemayresultinthevesselexplodingorblowingoffitscapwithdangerousforcecreatingflyingdebris.

Unpacking&StorageInstructions
IntendedUse
1. Checkallcontainersforleakageorbreakage.
Thisproductisintendedforresearchuseonly.Itisnot
2. Removethefrozencellsfromthedryicepackagingandimmediatelyplacethecellsatatemperature
intendedforanyanimalorhumantherapeuticor
below130C,preferablyinliquidnitrogenvapor,untilreadyforuse.
diagnosticuse.

HandlingProcedureforFrozenCells
CompleteGrowthMedium

ThebasemediumforthiscelllineisATCCformulated
Dulbecco'sModifiedEagle'sMedium,CatalogNo.30
2002.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:bovinecalf
serumtoafinalconcentrationof10%.
CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:3T3L1(ATCCCL173)
HandlingProcedureforFrozenCells
Toinsurethehighestlevelofviability,thawthevialandinitiatethecultureassoonaspossibleuponreceipt.If
uponarrival,continuedstorageofthefrozencultureisnecessary,itshouldbestoredinliquidnitrogenvapor
phaseandnotat
70C.Storageat70Cwillresultinlossofviability.
SAFETYPRECAUTION:ATCChighlyrecommendsthatprotectiveglovesandclothingalwaysbe
usedandafullfacemaskalwaysbewornwhenhandlingfrozenvials.Itisimportanttonotethat
somevialsleakwhensubmersedinliquidnitrogenandwillslowlyfillwithliquidnitrogen.Uponthawing,the
conversionoftheliquidnitrogenbacktoitsgasphasemayresultinthevesselexplodingorblowingoffits
capwithdangerousforcecreatingflyingdebris.
1.Thawthevialbygentleagitationina37Cwaterbath.Toreducethepossibilityofcontamination,keep
theOringandcapoutofthewater.Thawingshouldberapid(approximately2minutes).
2.Removethevialfromthewaterbathassoonasthecontentsarethawed,anddecontaminatebydipping
inorsprayingwith70%ethanol.Alloftheoperationsfromthispointonshouldbecarriedoutunderstrict
asepticconditions.
3.Transferthevialcontentstoacentrifugetubecontaining9.0mLcompleteculturemediumandspinat
approximately125xgfor5to7minutes.Discardsupernatant.
4.Resuspendthecellpelletwiththerecommendedcompletemediumanddispenseintoa25cm2culture
flask.Itisimportanttoavoidexcessivealkalinityofthemediumduringrecoveryofthecells.Itissuggested
that,priortotheadditionofthevialcontents,theculturevesselcontainingthecompletegrowthmediumbe
placedintotheincubatorforatleast15minutestoallowthemediumtoreachitsnormalpH(7.0to7.6).
5.Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif
usingthemediumdescribedonthisproductsheet.

HandlingProcedureforFlaskCultures

AmericanTypeCultureCollection HandlingProcedureforFlaskCultures
POBox1549
Theflaskwasseededwithcells(seespecificbatchinformation),grown,andcompletelyfilledwithmediumat
Manassas,VA20108USA
www.atcc.org
ATCCtopreventlossofcellsduringshipping.

800.638.6597or703.365.2700 1.Uponreceiptvisuallyexaminethecultureformacroscopicevidenceofanymicrobialcontamination.Using
Fax:703.365.2750 aninvertedmicroscope(preferablyequippedwithphasecontrastoptics),carefullycheckforanyevidence
Email:Tech@atcc.org
ofmicrobialcontamination.Alsochecktodetermineifthemajorityofcellsarestillattachedtothebottomof

Orcontactyourlocaldistributor theflaskduringshippingtheculturesaresometimeshandledroughlyandmanyofthecellsoftendetachand
becomesuspendedintheculturemedium(butarestillviable).
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 2.Ifthecellsarestillattached,asepticallyremoveallbut5to10mLoftheshippingmedium.The
shippingmediumcanbesavedforreuse.Incubatethecellsat37Cina5%CO2inairatmosphereuntilthey
arereadytobesubcultured.
3.Ifthecellsarenotattached,asepticallyremovetheentirecontentsoftheflaskandcentrifugeat125
xgfor5to10minutes.Removeshippingmediumandsave.Resuspendthepelletedcellsin10mlofthis
ProductSheet mediumandaddto25cm2flask.Incubateat37Cina5%CO2inairatmosphereuntilcellsarereadytobe
subcultured.
3T3L1(ATCCCL173)
SubculturingProcedure

PleasereadthisFIRST Protocol:Neverallowculturetobecomecompletelyconfluent.
1. Removeanddiscardculturemedium.
StorageTemp. 2. Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin0.53mMEDTAsolutiontoremovealltracesof
serumthatcontainstrypsininhibitor.
liquidnitrogen
3. Add2.0to3.0mLofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
vaporphase
untilcelllayerisdispersed(usuallywithin5to15minutes).
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthe
cellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.
4. Add6.0to8.0mlofcompletegrowthmediumandaspiratecellsbygentlypipetting.

BiosafetyLevel
5. Addappropriatealiquotsofthecellsuspensiontonewculturevessels.
1
Therecommendedinoculumis2to3X103cells/cm2.Subculturebeforeculturesbecome70to80%
confluentorwhencellsreach5to6X10(4)viablecells/cm2.CorningT75flasks(catalog#431464)
arerecommendedforsubculturingthisproduct.
IntendedUse 6. Incubateculturesat37C.
Interval:Everythreedays
Thisproductisintendedforresearchuseonly.Itisnot MediumRenewal:2to3timesperweek
intendedforanyanimalorhumantherapeuticor
diagnosticuse. CryopreservationMedium

CompleteGrowthMedium Completeculturemediumdescribedabovesupplementedwith5%(v/v)DMSO.CellculturetestedDMSOis
availableasATCCCatalogNo.4X.
ThebasemediumforthiscelllineisATCCformulated
Dulbecco'sModifiedEagle'sMedium,CatalogNo.30
Comments
2002.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:bovinecalf
Thecellsundergoapreadiposetoadiposelikeconversionastheyprogressfromarapidlydividingtoa
serumtoafinalconcentrationof10%.
confluentandcontactinhibitedstate.Ahighserumcontentinthemediumenhancesfataccumulation[PubMed
ID:4426090].
CitationofStrain
Testedandfoundnegativeforectromeliavirus(mousepox).
ThislineisalsodesignatedasATCCCCL92.1.ATCCCL173wasdepositedin1974withoutpassagenumber
Ifuseofthiscultureresultsinascientificpublication,it
informationfromthedepositor.
shouldbecitedinthatmanuscriptinthefollowing
manner:3T3L1(ATCCCL173)
References

Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:1

Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty

TheviabilityofATCCproductsiswarrantedfor30daysfromthedateofshipment,andisvalidonlyifthe
productisstoredandculturedaccordingtotheinformationincludedonthisproductinformationsheet.ATCC
AmericanTypeCultureCollection
liststhemediaformulationthathasbeenfoundtobeeffectiveforthisstrain.Whileother,unspecifiedmedia
POBox1549
Manassas,VA20108USA mayalsoproducesatisfactoryresults,achangeinmediaortheabsenceofanadditivefromtheATCC
www.atcc.org recommendedmediamayaffectrecovery,growthand/orfunctionofthisstrain.Ifanalternativemedium
formulationisused,theATCCwarrantyforviabilityisnolongervalid.
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org Disclaimers

Orcontactyourlocaldistributor
Thisproductisintendedforlaboratoryresearchpurposesonly.Itisnotintendedforuseinhumans.

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 WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproductsheet,
ATCCmakesnowarrantiesorrepresentationsastoitsaccuracy.Citationsfromscientificliteratureand
patentsareprovidedforinformationalpurposesonly.ATCCdoesnotwarrantthatsuchinformationhasbeen
confirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,anduse.ATCC
isnotliableforanydamagesorinjuriesarisingfromreceiptand/oruseofthisproduct.Whilereasonableeffort
ProductSheet
ismadetoinsureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
3T3L1(ATCCCL173) themisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org

PleasereadthisFIRST
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2015.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[02/25]
StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
1

IntendedUse

Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium

ThebasemediumforthiscelllineisATCCformulated
Dulbecco'sModifiedEagle'sMedium,CatalogNo.30
2002.Tomakethecompletegrowthmedium,addthe
followingcomponentstothebasemedium:bovinecalf
serumtoafinalconcentrationof10%.

CitationofStrain

Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:3T3L1(ATCCCL173)

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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