MatTek/EpiDermFT™/epidermft/1 Ea

Engineeredtoenableinvitroskinresearchinwhichfibroblast-keratinocytecellinteractionsareimportant,MatTek’sEpiDermFTsystemconsistsofnormal,humanepidermalkeratinocytes(NHEK)andnormal,humandermalfibroblasts(NHFB)culturedtoformamultilayeredmodelofthehumandermisandepidermis.Culturedattheair-liquidinterfaceineasy-to-handletissuecultureinserts,EpiDermFTattainslevelsofdifferentiationatthecuttingedgeoftissueengineeringtechnology.

EpiDermFTconsistsoforganizedkeratin5expressingbasalcells,involucrinandkeratin10expressingspinousandgranularlayers,andcornifiedepidermallayersanalogoustothosefoundinvivo.Thedermalcompartmentiscomposedofacollagenmatrixcontainingviablenormalhumandermalfibroblasts(NHDF).Theepidermalanddermallayersaremitoticallyandmetabolicallyactiveandexhibitinvivo-likemorphologicalandgrowthcharacteristicswhichareuniformandhighlyreproducIBLe.

Awell-developedbasementmembraneispresentatthedermal/epidermaljunction.Hemidesomosomes(›),laminalucida,laminadensa(→)andanchoringfibrilstructures(»)areevidentbytransmissionelectronmicroscopy.

EpiDermFTtissuesareusedacrossawiderangeofapplicationsincludingscientificefficacyandclaimssubstantiation.Duetoadryskinsurface,EpiDermFTcanbeusedtoevaluateBIOLOGicalresponsestotopicalapplicationsofformulationsoringredients.Simpleprotocolsandtheevaluationofearlymolecularendpointsallowresearchtoacquiredataindays,notweeksormonths.

Anti-Aging

Examinechangesinextracellularmatrixgeneandproteinexpressionandhistologicalstructuretodeterminetheanti-agingefficacyofcosmeticingredientsorfinalformulation.EpiDermFTisavailablefromarangeofadultdonors.

Anti-AgingApplicationNote

WoundHealing

UseEpiDermFTtoevaluatetherapeuticswiththewoundhealingassaykit.Woundhealingassaykitsinclude24pre-woundedtissuesandareavailablewithepidermalonlywoundsorfullthicknesswoundsoftheepidermisanddermis.

RecentWoundHealingPoster

SkinHydration

UseEpiDermFTtoassesstheefficacyoftopicallyappliedmoisturizersbymeasuringtheelectricalimpedanceoftheepidermis.Amenabletocreams,gel,lotionsorotherformulations,theSkinHydrationassayallowsfordataanalysiswithin24hours.

SkinHydrationApplicationNote

UVProtection

Analyzecyclopyrimidinedimerformation,inflammatorymediatorreleaseandoxidativeendpointstoevaluatetheUVprotectiveefficacyofcompoundsorformulations.

UVProtection ApplicationNote

BrowseourreferencelibrarytoseehowourEpiDermFT tissuehasbeenusedintheseareasofstudy.

Tissue

Kit: AstandardEpiDermFTkit(EFT-400)consistsof24tissues.(Tissue“kits”containtissues,asmallamountofculturemedium,andplasticware;contactMatTekforspecifickitcontents)

Substrate: CostarSnapwell™singlewelltissuecultureplateinserts areused.Poresize=0.4μm,Diameter=1.2cm.Surfacearea=1.0cm²

Culture: Air-liquidinterface

Histology: 8-12celllayersplusstratumcorneum(basal,spinous,andgranularlayers)

Lotnumbers: Tissuelotsproducedeachweekareassignedaspecificlotnumber.Aletterofthealphabetisappendedtotheendofthelotnumbertodifferentiatebetweenindividualkitswithinagivenlotoftissues.Alltissuekitswithinalotareidenticalinregardstocells,medium,handling,cultureconditions,etc.

Shipment: At4°Conmedium-supplemented,agarosegelsin6-wellplates

Shipmentday: EveryMonday.ShipmentonThursdayalsopossibleuponspecialrequest

Delivery: TuesdaymorningviaFedExpriorityservice(US).OutsideUS:Tuesday-Thursdaydependingonlocation

Shelflife: Includingtimeintransit,tissuesmaybestoredat4°Cforupto6dayspriortouse.However,extendedstorageperiodsarenotrecommendedunlessnecessary.Inaddition,thebestreproducibilitywillbeobtainediftissuesareusedconsistentlyonthesameday,e.g.Tuesdayafternoonorfollowingovernightstorageat4°C(Wednesdaymorning)

Lengthofexperiments: Culturescanbecontinuedforupto2weekswithgoodretentionofnormalepidermalmorphology.Culturesmustbefedeveryotherdaywith5.0mlofEFT-400medium

Cells

Type: Normalhumanepidermalkeratinocytes(NHEK);Normalhumandermalfibroblasts(NHDF)

Geneticmake-up: Singledonor

Derivedfrom: Neonatal-foreskintissue(NHEK);Neonatalskin(NHDF)

Alternatives: NHEKfromadultbreastskin;NHDFfromadultskin

Screenedfor: HIV,Hepatitis-B,Hepatitis-C,mycoplasma

Medium

Basemedium: Dulbecco’sModifiedEagle’sMedium(DMEM)

Growthfactors/hormones: Epidermalgrowthfactor,insulin,hydrocortisoneandotherproprietarystimulatorsofepidermaldifferentiation

Serum: None

Antibiotics: Gentamicin5µg/ml(10%ofnormalgentamicinlevel)

Anti-fungalagent: AmphotericinB0.25µg/ml

pHIndicator: Phenolred

Otheradditives: Lipidprecursorsusedtoenhanceepidermalbarrierformation(proprietary)

Alternatives: Phenolred-free(EFT-400-PRF),antibiotic-free(EFT-400-ABF),anti-fungal-free(EFT-400-AFF),orhydrocortisone-freemediumandtissue(EFT-400-HCF)areavailable.Agentsareremovedatleast3dayspriortoshipment

Assay/Maintenancemedium: EFT-400-ASYisutilizedforassays;EFT-400-MMisusedfortermmaintenanceoftheEFT-400tissues

QualityControlandSterility

Visualinspection: Alltissuesarevisuallyinspectedandifphysicalimperfectionsarenoted,tissuesarerejectedforshipment

End-usetesting: Tissuesareexposedto1%TritonX-100for8,12,16and24hours.ThetimeofexposurerequiredtoreducethetissueviABIlity(ET-50)usingtheMTTviabilityassayisdetermined(SeeMatTekEpiDermFTuseprotocol)foreachlotoftissue.ET-50’sgenerallyfallwithintherangeof6.5-9.5hours.ET-50’sincustomers’labmaydifferslightlyfromtheMatTekresults

Sterility: Allmediausedthroughouttheproductionprocessischeckedforsterility.Maintenancemediumisincubatedwithandwithoutantibioticsfor1weekandcheckedforsterility.Theagarosegelfromthe24-wellplateusedforshippingisalsoincubatedfor1weekandcheckedforanysignofcontamination

Screeningforpathogens: AllcellsarescreenedandarenegativeforHIV,hepatitisBandhepatitisCusingPCR.However,noknowntestmethodcanoffercompleteassurancethatthecellsarepathogenfree.Thus,theseproductsandallhumanderivedproductsshouldbehandledatBSL-2levels(biosafetylevel2)orhigherasrecommendedintheCDC-NIHmanual,“Biosafetyinmicrobiologicalandbiomedicallaboratories,”1998.Forfurtherassistance,pleasecontactyoursiteSafetyOfficerorMatTektechnicalservice

Notificationoflotfailure: IfatissuelotfailsourQCorsterilitytesting,thecustomerwillbenotifiedandthetissueswillbereplacedwithoutchargewhenappropriate.BecauseourQCandsterilitytestingisdonepost-shipment,notificationwillbemadeassoonaspossible(Undernormalcircumstances,ET-50failureswillbenotifiedbyWednesday5p.m.;sterilityfailureswillbenotifiedwithin8daysofshipment)