MatTek/EpiDerm™/epiderm/1 Ea

AlsoknowngenericallyasaReconstructedHumanEpidermis(RHE),EpiDermisaready-to-use,highlydifferentiated3Dtissuemodelconsistingofnormal,human-derivedepidermalkeratinocytes(NHEK)culturedonspeciallypreparedtissuecultureinserts.

Culturedattheair-liquidinterface(ALI),EpiDermallowsfortheevaluationoftopicallyappliedcompounds,chemicals,cosmetic/personalcareproductingredientsandfinalformulations.Withourstraightforwardprotocolsand20+yearsofdata,EpiDermisthegoldstandardforabroadrangeofhighlypredictiveinvitroapplications.

EpiDermexhibitshumanepidermaltissuestructureandcellularmorphologywithgreateruniformityandreproducibility.It’s3Dstructureconsistingoforganizedandproliferativebasalcells,spinousandgranularlayers,andcornifiedepidermallayersaremitoticallyandmetabolicallyactive.

TheEpiDerm3Dhumantissuemodelisusedacrossadiverserangeofapplicationsincludingsafetyandriskassessment,andBIOLOGicalefficacy.WithAir-liquidInterface(ALI)culturetechnology,EpiDermcanbeusedtoevaluatebiologicalresponsestotopicalapplicationsofformulationsoringredients.Simpleprotocolsandtheevaluationofearlycellularendpointsallowresearchtoacquiredataindays,notweeksormonths.

SkinIrritation(OECDTG439)

UseEpiDermtodeterminetheskinirritationpotentialofliquids,solids,gels,lotions,ointmentsorcreams.TissueviABIlityresultsareacquiredwithin3days.

SkinIrritationTestProtocol

SkinCorrosion(OECDTG431)

Determinetheskincorrosionpotentialoftestmaterialsinasingleday.

SkinCorrosionProtocol

SkinHydration

MeasuretheelectricalimpedanceofEpiDermtoassesstheefficacyoftopicallyappliedmoisturizersincludingcreams,gelsandlotions.TheSkinHydrationprotocolallowsfordataanalysiswithin24hours.

SkinHydrationApplicationNote

DermalDrugDelivery

UtilizeexistingtransdermalpermeationequipmentorMatTek’ssimplesingleinsertpermeationdevicestoassessAPIpermeationandflux.DeterminerelativesafetywiththeEpiDermMTTET-50assay.

PercutaneousAbsorptionProtocol

Phototoxicity

IdentifyphototoxiceffectsoftopicalorsystemictestsubstancesusingtheEpiDermPhototoxicityAssay(pre-validated)

PhototoxicityProtocol

DermalGenotoxicity

IdentifygenotoxiceffectsoftopicalorsystemictestsubstancesusingEpiDermGenotoxicityAssays(pre-validated)

MicronucleusAssayProtocol

CometAssayProtocol

EpidermalDifferentiation

Evaluateactivecompoundsforeffectsonepidermaldifferentiationusingtheunder-developedEpiDermtissuemodel

BrowseourreferencelibrarytoseehowourEpiDermtissuehasbeenusedintheseareasofstudy.

Tissue

Kit:AstandardEpiDermkit(EPI-200)consistsof24tissues.(Tissue“kits”containtissues,asmallamountofculturemedium,andplasticware;contactMatTekforspecifickitcontents)

Formats:9mm&22mmindividualinsertsand96-wellHTSplates–tissueculturesubstrateischemicallymodifieswithaporesizeof0.4μm

Culture:Air-liquidinterface

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

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

Shipment:At4°Conmedium-supplemented,agarosegels

Shipmentday:EveryMonday.ShipmentonThursdayalsopossIBLeuponspecialrequest

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

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

Lengthofexperiments:Culturescanbecontinuedforupto3weekswithgoodretentionofnormalepidermalmorphology.Culturesmustbefedeveryotherdaywith5.0mloflonglifemaintenancemedium(EPI-100-LLMM),standardmaintenancemedium(EPI-100-MM),ornewmaintenancemedium(EPI-100-NMM).Extendedcultureexperimentsrequiretheuseofculturestands(MEL-STND;seephoto)orwashers(EPI-WSHR)in6-wellplatestoallowforculturingin5.0mlofculturemedium

Cells

Type:Normalhumanepidermalkeratinocytes(NHEK)

Geneticmake-up:Singledonor

Derivedfrom:Neonatal-foreskintissue(NHEK)

Alternatives:NHEKfromadultbreastskin

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(EPI-200-PRF),antibiotic-free(EPI-200-ABF),anti-fungal-free(EPI-200-AFF),orhydrocortisone-freemediumandtissue(EPI-200-HCF)areavailable.Agentsareremovedatleast3dayspriortoshipment.PleasediscussyourspecificmediaformulationandtissueneedswithMatTekscientificpersonnelbeforeplacingyourorder.

Assay/Maintenancemedium:Threemaintenancemediaareavailable.EPI-100-MMisequivalenttotheassaymediumusedforshorttermtoxicologicaltesting.LongLifeMaintenanceMedium,EPI-100-LLMM,containsα-MSHandβ-FGF.NewMaintenanceMedium,EPI-100-NMM,containsKeratinocyteGrowthFactor(KGF),butdoesnotcontainα-MSHandβ-FGF.Althoughallmediawillresultingoodretentionofepidermalmorphologyforupto3weeks(5.0mlchangedeveryotherday),useofEPI-100-NMMwillgivetheoptimalhistologyforlongtermstudies.

QualityControlandSterility

Visualinspection:Alltissuesarevisuallyinspectedandifphysicalimperfectionsarenoted,tissuesarerejectedforshipment

End-usetesting:Tissuesareexposedto1%TritonX-100for4,6,8and12.5hours.Thetimeofexposurerequiredtoreducethetissueviability(ET-50)usingtheMTTviabilityassayisdetermined(SeeMatTekEpiDermMTTET-50Protocol)foreachlotoftissue.ET-50’smustfallwithintherangeofthe1996EpiDermdatabaseof4.77–8.72hours.ET-50’sincustomers’labsmaydifferslightlyfromtheMatTekresults.

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)