Scribd
Upload
202 views3 pages

Ect1/E6E7 (ATCC CRL 2614) : Product Sheet

This document provides product information for the Ect1/E6E7 cell line. It describes the organism, tissue, cell type, morphology, growth properties, storage conditions, biosafety level, intended use, complete growth medium, citations, handling procedures, and references. The document contains detailed instructions for culturing, subculturing, cryopreserving, and handling 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
202 views3 pages

Ect1/E6E7 (ATCC CRL 2614) : Product Sheet

This document provides product information for the Ect1/E6E7 cell line. It describes the organism, tissue, cell type, morphology, growth properties, storage conditions, biosafety level, intended use, complete growth medium, citations, handling procedures, and references. The document contains detailed instructions for culturing, subculturing, cryopreserving, and handling 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

Ect1/E6E7(ATCCCRL
2614)

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

BatchSpecificInformation
PleasereadthisFIRST
RefertotheCertificateofAnalysisforbatchspecifictestresults.

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

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

Unpacking&StorageInstructions
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:Ect1/E6E7(ATCCCRL2614)

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

HandlingProcedureforFrozenCells
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

Ect1/E6E7(ATCCCRL
2614)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

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:Ect1/E6E7(ATCCCRL2614)

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:1:4to1:6
MediumRenewal:Every2to3days.
Note:FormoreinformationonenzymaticdissociationandsubculturingofcelllinesconsultChapter10in
CultureofAnimalCells,[Link],3rdedition,publishedbyAlan
[Link],N.Y.,1994

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

Orcontactyourlocaldistributor

Page2of3

[Link].

BiosafetyLevel:2
[Link]
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty

ProductSheet

Ect1/E6E7(ATCCCRL
2614)

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

Disclaimers
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vaporphase

BiosafetyLevel
2

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:Ect1/E6E7(ATCCCRL2614)

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

Orcontactyourlocaldistributor

Page3of3

[Link].
WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproductsheet,
[Link]
[Link]
confirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,[Link]
isnotliableforanydamagesorinjuriesarisingfromreceiptand/[Link]
ismadetoensureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
[Link]
[Link].
[Link].[01/14]

You might also like