Description

ProductSheet

BEAS2B(ATCCCRL
9609)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
Thebasemediumforthiscellline(BEBM)alongwithall
theadditivescanbeobtainedfromLonza/Clonetics
Corporationasakit:BEGM,KitCatalogNo.CC3170.
ATCCdoesnotusetheGA1000(gentamycin
amphotericinBmix)providedwiththeBEGMkit.Note:
Donotfiltercompletemedium.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:BEAS2B(ATCCCRL9609)

Organism:Homosapiens,human
Tissue:lung/bronchus
Disease:normal
CellType:epithelial(virustransformed)
Morphology:epithelial
GrowthProperties:adherent
DNAProfile:
Amelogenin:XY
CSF1PO:9,12
D13S317:13
D16S539:12
D5S818:12,13
D7S820:10,13
THO1:7,9.3
TPOX:6,11
vWA:17,18

BatchSpecificInformation
RefertotheCertificateofAnalysisforbatchspecifictestresults.

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

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

HandlingProcedureforFrozenCells
Toinsurethehighestlevelofviability,thawthevialandinitiatethecultureassoonaspossibleuponreceipt.If
uponarrival,continuedstorageofthefrozencultureisnecessary,itshouldbestoredinliquidnitrogenvapor
phaseandnotat70C.Storageat70Cwillresultinlossofviability.
Note:Thecultureflasksusedshouldbeprecoatedwithamixtureof0.01mg/mLfibronectin,0.03mg/mL
bovinecollagentypeIand0.01mg/mLbovineserumalbumindissolvedinBEBMmedium(seereferences:U.S.
Pat.4,885,238andLechner,J.F.andLaVeck,M.A.Aserumfreemethodforculturingnormalhumanbronchial
epithelialcellsatclonaldensity.J.TissueCultureMethods9:4348,1985).
1. Thawthevialbygentleagitationina37Cwaterbath.Toreducethepossibilityofcontamination,keep
theOringandcapoutofthewater.Thawingshouldberapid(approximately2minutes).
2. Removethevialfromthewaterbathassoonasthecontentsarethawed,anddecontaminateby
dippinginorsprayingwith70%ethanol.Alloftheoperationsfromthispointonshouldbecarriedout
understrictasepticconditions.
3. Transferthevialcontentstoacentrifugetubecontaining9.0mLcompleteculturemediumand
centrifugethecellsuspensionatapproximately125xgfor5to10minutes.Discardthesupernatant.
4. Resuspendthecellpelletinthecompleteculturemediumatthedilutionratiorecommendedinthe
specificbatchinformationanddispenseintoaprecoatedT25cm2cultureflask.
5. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif

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

Orcontactyourlocaldistributor

Page1of3

usingthemediumdescribedonthisproductsheet.

HandlingProcedureforFlaskCultures
Theflaskwasseededwithcells(seespecificbatchinformation),grown,andcompletelyfilledwithmediumat
ATCCtopreventlossofcellsduringshipping.
1. Uponreceiptvisuallyexaminethecultureformacroscopicevidenceofanymicrobialcontamination.
Usinganinvertedmicroscope(preferablyequippedwithphasecontrastoptics),carefullycheckfor
anyevidenceofmicrobialcontamination.Alsochecktodetermineifthemajorityofcellsarestill

attachedtothebottomoftheflaskduringshippingtheculturesaresometimeshandledroughlyand
manyofthecellsoftendetachandbecomesuspendedintheculturemedium(butarestillviable).
2. Ifthecellsarestillattached,asepticallyremoveallbut5to10mLoftheshippingmedium.The
shippingmediumcanbesavedforreuse.Incubatethecellsat37Cina5%CO2inairatmosphere

ProductSheet

BEAS2B(ATCCCRL
9609)

untiltheyarereadytobesubcultured.
3. Ifthecellsarenotattached,asepticallyremovetheentirecontentsoftheflaskandcentrifugeat
125xgfor5to10minutes.Removeshippingmediumandsave.Resuspendthepelletedcellsin10
mLofthismediumandaddtoprecoated25cm2flask.Incubateat37Cina5%CO2inair
atmosphereuntilcellsarereadytobesubcultured.

SubculturingProcedure
PleasereadthisFIRST

Thesecellsshouldbesubculturedbeforereachingconfluencesinceconfluentculturesrapidlyundergo
squamousterminaldifferentiation.Volumesusedinthisprotocolarefor75cm 2flaskproportionallyreduceor

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
Thebasemediumforthiscellline(BEBM)alongwithall
theadditivescanbeobtainedfromLonza/Clonetics
Corporationasakit:BEGM,KitCatalogNo.CC3170.
ATCCdoesnotusetheGA1000(gentamycin
amphotericinBmix)providedwiththeBEGMkit.Note:
Donotfiltercompletemedium.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:BEAS2B(ATCCCRL9609)

increaseamountofdissociationmediumforculturevesselsofothersizes.CorningT75flasks(catalog
#430641)arerecommendedforsubculturingthisproduct.
1. Removeanddiscardculturemedium.
2. Add2.0to3.0mLof0.25%Trypsin0.53mMEDTAsolutioncontaining0.5%polyvinylpyrrolidone
(PVP)toflaskandobservecellsunderaninvertedmicroscopeuntilcelllayerisdispersed(usually
with5to10minutes).
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaiting
forthecellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitate
dispersal.
3. Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.
4. Transfercellsuspensiontocentrifugetubeandspinatapproximately125xgfor5to10minutes
5. Discardsupernatantandresuspendcellsinfreshgrowthmedium.Inoculatenewflasksat1500to
3000cellspercm2.Thecultureflasksusedshouldbeprecoatedwithamixtureof0.01mg/ml
fibronectin,0.03mg/mlbovinecollagentypeIand0.01mg/mLbovineserumalbumindissolvedin
BEBM.
6. Placecultureflasksinincubatorsat37C.
Note:FormoreinformationonenzymaticdissociationandsubculturingofcelllinesconsultChapter10in
CultureofAnimalCells,amanualofBasicTechniquebyR.IanFreshney,3rdedition,publishedby
AlanR.Liss,N.Y.,1994.
Interval:Subculturedbeforereachingconfluence.
MediumRenewal:Every2to3days
FlaskCoating
1. Prepareamixtureof0.01mg/mLfibronectin,0.03mg/mLbovinecollagentypeIand0.01mg/mLbovine
serumalbumin(BSA)dissolvedinculturemedium.StoreprepreparedCoatingSolutionat4Cincold
roomforupto3months.
2. Foragrowthareaof75cm2,add4.5mLofthefibronectin/collagen/BSAsolutionandrockgentlyto
coattheentiresurface.
3. Incubatethefreshlycoatedvessel(s)ina37Cincubatorovernight(itispreferabletousetissue
culturevesselswithtightened,plugsealcapstopreventevaporationduringthecoatingprocess).
4. Storecoatedflaskswithsolutionatroomtemperature,lightprotected,upto1month.Suctionoff
solutionbeforeplatingcells.

CryopreservationMedium
CryoprotectantMedium
Completegrowthmediumplus1%PVPand7.5%DMSO.
CellculturetestedDMSOisavailableasATCCCatalogNo.4X.

Comments
ThecellsstainpositivelyforkeratinsandSV40Tantigen.
AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
www.atcc.org
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@atcc.org

Orcontactyourlocaldistributor

References
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:2
Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin

Page2of3

thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty

ProductSheet

BEAS2B(ATCCCRL
9609)

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

PleasereadthisFIRST

Disclaimers

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
Thebasemediumforthiscellline(BEBM)alongwithall
theadditivescanbeobtainedfromLonza/Clonetics
Corporationasakit:BEGM,KitCatalogNo.CC3170.
ATCCdoesnotusetheGA1000(gentamycin
amphotericinBmix)providedwiththeBEGMkit.Note:
Donotfiltercompletemedium.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:BEAS2B(ATCCCRL9609)

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
ismadetoinsureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2014.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[08/26]