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Description

ProductSheet

HT29(ATCCHTB38)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vapor
temperature

BiosafetyLevel
1

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
McCoy's5aMediumModified,CatalogNo.302007.To
makethecompletegrowthmedium,addthefollowing
componentstothebasemedium:fetalbovineserumto
afinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HT29(ATCCHTB38)

Organism:Homosapiens,human
Tissue:
colon
Disease:colorectaladenocarcinoma
Age:44yearsadult
Gender:female
Morphology:epithelial
GrowthProperties:adherent
Isoenzymes:
AK1,1
ESD,1
G6PD,B
GLOI,12
Me2,1
PGM1,12
PGM3,12
DNAProfile:
Amelogenin:X
CSF1PO:11,12
D13S317:11,12
D16S539:11,12
D5S818:11,12
D7S820:10
THO1:6,9
TPOX:8,9
vWA:17,19
CytogeneticAnalysis:modalnumber=71range=68to72.
Thestemlinechromosomenumberishypertriploidwiththe2Scomponentoccurringat2.4%.Seventeen
markerchromosomesarefoundinmostmetaphases,generallyinsinglecopyperchromosome.Themarker
designationsare:M1p(=t(3p?)withadeletedshortarm),t(7q?),t(10q?),i(13q),19q+aM6,?t(8q9q),?
Xp,M9,6q+,t(13?)a,t(13?)b,19q+b,M14,M15,15p+,andXq.Chromosome13isnullisomicand
chromosomes8and14aregenerallymonosomic.NoYchromosomewasdetectedbyQMbandanalysis.

BatchSpecificInformation
RefertotheCertificateofAnalysisforbatchspecifictestresults.

SAFETYPRECAUTION
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. Transferthevialcontentstoacentrifugetubecontaining9.0mLcompleteculturemedium.andspinat
approximately125xgfor5to7minutes.

4. Resuspendcellpelletwiththerecommendedcompletemedium(seethespecificbatchinformationfor

ProductSheet

HT29(ATCCHTB38)

theculturerecommendeddilutionratio).anddispenseintoa25cm2cultureflask.Itisimportantto
avoidexcessivealkalinityofthemediumduringrecoveryofthecells.Itissuggestedthat,priortothe
additionofthevialcontents,theculturevesselcontainingthecompletegrowthmediumbeplacedinto
theincubatorforatleast15minutestoallowthemediumtoreachitsnormalpH(7.0to7.6).pH(7.0to
7.6).
5. Incubatethecultureat37Cinasuitableincubator.A5%CO2inairatmosphereisrecommendedif
usingthemediumdescribedonthisproductsheet.

PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vapor
temperature

BiosafetyLevel
1

IntendedUse

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.

Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
McCoy's5aMediumModified,CatalogNo.302007.To
makethecompletegrowthmedium,addthefollowing
componentstothebasemedium:fetalbovineserumto
afinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HT29(ATCCHTB38)

SubculturingProcedure
1. Removeanddiscardculturemedium.
2. Brieflyrinsethecelllayerwith0.25%(w/v)Trypsin0.53mMEDTAsolutiontoremovealltracesof
serumwhichcontainstrypsininhibitor.
3. Add2.0to3.0mLofTrypsinEDTAsolutiontoflaskandobservecellsunderaninvertedmicroscope
untilcelllayerisdispersed(usuallywithin5to15minutes).CorningT75flasks(catalog#430641)
arerecommendedforsubculturingthisproduct.
Note:Toavoidclumpingdonotagitatethecellsbyhittingorshakingtheflaskwhilewaitingforthe
cellstodetach.Cellsthataredifficulttodetachmaybeplacedat37Ctofacilitatedispersal.
4. Add6.0to8.0mLofcompletegrowthmediumandaspiratecellsbygentlypipetting.
5. Addappropriatealiquotsofthecellsuspensiontonewculturevessels.
6. Incubateculturesat37C.
SubcultivationRatio:Asubcultivationratioof1:3to1:8isrecommended
MediumRenewal:2to3timesperweek

CryopreservationMedium
Completegrowthmediumdescribedabovesupplementedwith5%(v/v)DMSO.
CellculturetestedDMSOisavailableasATCCCatalogNo.4X.

Comments

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

Orcontactyourlocaldistributor

Page2of3

UltrastructuralfeaturesreportedforHT29cellsincludemicrovilli,microfilaments,largevacuolated
mitochondriawithdarkgranules,smoothandroughendoplasmicreticulumwithfreeribosomes,lipiddroplets,
fewprimaryandmanysecondarylysosomes.
Thecellsexpressurokinasereceptors,butdonothavedetectableplasminogenactivatoractivity[PubMedID:
8381394].HT29cellsarenegativeforCD4,butthereiscellsurfaceexpressionofgalactoseceramide(a
possiblealternativereceptorforHIV).
Thelineispositiveforexpressionofcmyc,Kras,Hras,Nras,Myb,sisandfosoncogenes.Thep53
antigenisoverproduced,andthereisaG>Amutationincodon273ofthep53generesultinginanArg>
Hissubstitution.Nmyconcogeneexpressionwasnotdetected.
ThereisaG>Amutationincodon273ofthep53generesultinginanArg>Hissubstitution.

References
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.

BiosafetyLevel:1

ProductSheet

HT29(ATCCHTB38)
PleasereadthisFIRST

StorageTemp.
liquidnitrogen
vapor
temperature

Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.Laboratorysafetyisdiscussedin
thecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.
DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
forHealth.

ATCCWarranty
ATCCproductsarewarrantedfor30daysfromthedateofshipment,andthiswarrantyisvalidonlyifthe
productisstoredandhandledaccordingtotheinformationincludedonthisproductinformationsheet.Ifthe
ATCCproductisalivingcellormicroorganism,ATCCliststhemediaformulationthathasbeenfoundtobe
effectiveforthisproduct.Whileother,unspecifiedmediamayalsoproducesatisfactoryresults,achangein
mediaortheabsenceofanadditivefromtheATCCrecommendedmediamayaffectrecovery,growthand/or
functionofthisproduct.Ifanalternativemediumformulationisused,theATCCwarrantyforviabilityisno
longervalid.

Disclaimers

BiosafetyLevel
1

IntendedUse
Thisproductisintendedforresearchuseonly.Itisnot
intendedforanyanimalorhumantherapeuticor
diagnosticuse.

CompleteGrowthMedium
ThebasemediumforthiscelllineisATCCformulated
McCoy's5aMediumModified,CatalogNo.302007.To
makethecompletegrowthmedium,addthefollowing
componentstothebasemedium:fetalbovineserumto
afinalconcentrationof10%.

CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,it
shouldbecitedinthatmanuscriptinthefollowing
manner:HT29(ATCCHTB38)

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
ismadetoinsureauthenticityandreliabilityofmaterialsondeposit,ATCCisnotliablefordamagesarisingfrom
themisidentificationormisrepresentationofsuchmaterials.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthis
product.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
ATCC2015.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[06/08]

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