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End1/E6E7 (ATCC CRL 2615) : Product Sheet

This document provides product information for the End1/E6E7 cell line, including descriptions of storage conditions, intended use, handling procedures, subculturing instructions, and safety precautions. It also includes background information on the cell line.

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109 views3 pages

End1/E6E7 (ATCC CRL 2615) : Product Sheet

This document provides product information for the End1/E6E7 cell line, including descriptions of storage conditions, intended use, handling procedures, subculturing instructions, and safety precautions. It also includes background information on the cell line.

Uploaded by

bogobon
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

Description

ProductSheet

End1/E6E7(ATCCCRL
2615)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

Organism:Homosapiens,human
Tissue:endocervixcervix
Disease:Endometriosis
CellType:epithelialHPV16E6/E7transformed
Age:43years
Gender:female
Morphology:epithelial
GrowthProperties:adherent

BatchSpecificInformation
RefertotheCertificateofAnalysisforbatchspecifictestresults.

SAFETYPRECAUTION
ATCChighlyrecommendsthatprotectiveglovesandclothingalwaysbeusedandafullfacemaskalwaysbe
[Link]
[Link],theconversionoftheliquidnitrogenbacktoitsgas
phasemayresultinthevesselexplodingorblowingoffitscapwithdangerousforcecreatingflyingdebris.

Unpacking&StorageInstructions
IntendedUse
[Link]
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

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

HandlingProcedureforFrozenCells
CompleteGrowthMedium
KeratinocyteSerumFreemedium(GIBCOBRL17005
042)with0.1ng/mlhumanrecombinantEGF,0.05
mg/mlbovinepituitaryextract,andadditionalcalcium
chloride44.1mg/L(finalconcentration0.4mM)

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:End1/E6E7(ATCCCRL2615)

Toinsurethehighestlevelofviability,[Link]
uponarrival,continuedstorageofthefrozencultureisnecessary,itshouldbestoredinliquidnitrogenvapor
phaseandnotat70C.Storageat70Cwillresultinlossofviability.
1. [Link],keep
[Link](approximately2minutes).
2. Removethevialfromthewaterbathassoonasthecontentsarethawed,anddecontaminateby
dippinginorsprayingwith70%[Link]
understrictasepticconditions.
3. [Link]
[Link]
[Link]
appropriateamountoffreshgrowthmedium.
4. [Link]
[Link],priortotheadditionofthevialcontents,the
culturevesselcontainingthegrowthmediumbeplacedintotheincubatorforatleast15minutesto
allowthemediumtoreachitsnormalpH(7.0to7.6).
5. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif
usingthemediumdescribedonthisproductsheet.

HandlingProcedureforFlaskCultures
Theflaskwasseededwithcells(seespecificbatchinformation),grown,andcompletelyfilledwithmediumat
ATCCtopreventlossofcellsduringshipping.
AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
[Link]
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@[Link]

Orcontactyourlocaldistributor

Page1of3

1. Uponreceipt,visuallyexaminethecultureformacroscopicevidenceofanymicrobialcontamination.
Usinganinvertedmicroscope(preferablyequippedwithphasecontrastoptics),carefullycheckfor
[Link],checktodetermineifthemajorityofcellsarestill
attachedtothebottomoftheflaskduringshippingtheculturesaresometimeshandledroughlyand
manyofthecellsoftendetachandbecomesuspendedintheculturemedium(butarestillviable).
2. Ifthecellsarestillattached,[Link]
shippingmediumcanbesavedforreuse.Incubatethecellsat37Cina5%CO2inairatmosphere
untiltheyarereadytobesubcultured.
3. Ifthecellsarenotattached,asepticallyremovetheentirecontentsoftheflaskandcentrifugeat

[Link].Resuspendthepelletedcellsin10
mLofthismediumandaddto25cm2flask.Incubateat37Cina5%CO2inairatmosphereuntilcells
arereadytobesubcultured.

SubculturingProcedure

ProductSheet

End1/E6E7(ATCCCRL
2615)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

IntendedUse
[Link]
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

Volumesusedinthisprotocolarefor75cm2flaskproportionallyreduceorincreaseamountofdissociation
mediumforculturevesselsofothersizes.
Note:Thecellsshouldnotbeallowedtobecomeconfluent,subcultureat60to90%ofconfluence.
1. Removeanddiscardculturemedium.
2. Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin0.03%(w/v)EDTAsolutiontoremovealltraces
ofserumthatcontainstrypsininhibitor.
3. Add2.0to3.0mlofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
untilcelllayerisdispersed(usuallywithin5to15minutes).
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthe
cellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.
4. Neutralizethetrypsinbyadding6.0to8.0mLofa1:1mixtureofDulbecco'smodifiedEagle'smedium
andHam'sF12mediumcontaining10%[Link].
5. ToremovetrypsinEDTAsolution,transfercellsuspensiontocentrifugetubeandspinat
approximately125xgfor5to10minutes.
6. [Link]
aliquotsofcellsuspensiontonewculturevessels.
7. Placeculturevesselsinincubatorsat37C.Cellswillnotattachwellfor24hoursaftersubculture.
SubcultivationRatio:1:3to1:5
MediumRenewal:Every2to3days
Note:FormoreinformationonenzymaticdissociationandsubculturingofcelllinesconsultChapter10in
CultureofAnimalCells,[Link],3rdedition,publishedbyAlan
[Link],N.Y.,1994

CompleteGrowthMedium

CryopreservationMedium
KeratinocyteSerumFreemedium(GIBCOBRL17005
042)with0.1ng/mlhumanrecombinantEGF,0.05
mg/mlbovinepituitaryextract,andadditionalcalcium
chloride44.1mg/L(finalconcentration0.4mM)

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:End1/E6E7(ATCCCRL2615)

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
[Link]
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@[Link]

Orcontactyourlocaldistributor

Page2of3

A1:1mixtureofDulbecco'smodifiedEagle'smediumandHam'sF12medium,85%fetalbovineserum,10%
DMSO,5%.
CellculturetestedDMSOisavailableasATCCCatalogNo.4X.

Comments
Theendocervicalcelllineexpressescharacteristicsofsimplecolumnarepithelium,whereastheectocervical
andvaginalcelllinesexpresscharacteristicsofstratifiedsquamousnonkeratinizingepithelia.
Withoutstimulation,allthreecelllinesproducemacrophagecolonystimulatingfactor(MCSF),transforming
growthfactorbeta1,interleukin8(IL8),prostaglandinE2,thesecretoryleukoproteinaseinhibitor,andthe
polymericimmunoglobulinreceptor.
Theendocervicalcellline(End1/E6E7),butnottheothers,alsoproducethelymphopoieticcytokinesIL6,IL7,
andconsistentlydetectablelevelsofthechemokineknownas"regulateduponactivation,normalTcell
expressedandsecreted"(RANTES).
Stimulationwithinterferongammaandtumornecrosisfactoralpha(TNFalpha)inducesorsignificantlyup
regulatesexpressionofseveralofthecytokinesandchemokinesaswellasmajorhistocompatibilitycomplex
(MHC)classIIantigensinthelines.
Piliated,butnotnonpiliated,NeisseriagonorrhoeastrainF62variantsactivelyinvadetheseepithelialcelllines.
Invasionofthesecellsbygreenfluorescentproteinexpressinggonococciischaracterizedbycolocalization
ofgonococciwithFactin.

References
[Link].

BiosafetyLevel:2
[Link]
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.

DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty

ProductSheet

End1/E6E7(ATCCCRL
2615)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

TheviabilityofATCCproductsiswarrantedfor30daysfromthedateofshipment,andisvalidonlyifthe
[Link]
[Link],unspecifiedmedia
mayalsoproducesatisfactoryresults,achangeinmediaortheabsenceofanadditivefromtheATCC
recommendedmediamayaffectrecovery,growthand/[Link]
formulationisused,theATCCwarrantyforviabilityisnolongervalid.

Disclaimers
[Link].
WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproductsheet,
[Link]
[Link]
confirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,[Link]
isnotliableforanydamagesorinjuriesarisingfromreceiptand/[Link]
ismadetoensureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
[Link]

IntendedUse
[Link]
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
KeratinocyteSerumFreemedium(GIBCOBRL17005
042)with0.1ng/mlhumanrecombinantEGF,0.05
mg/mlbovinepituitaryextract,andadditionalcalcium
chloride44.1mg/L(finalconcentration0.4mM)

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:End1/E6E7(ATCCCRL2615)

AmericanTypeCultureCollection
POBox1549
Manassas,VA20108USA
[Link]
800.638.6597or703.365.2700
Fax:703.365.2750
Email:Tech@[Link]

Orcontactyourlocaldistributor

Page3of3

[Link].
[Link].[06/06]

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