MatTek/EpiOcular™/epiocular/1 Ea

EpiOculareyeirritationassayshavebeenincorporatedintotoxicologylaboratoriesacrosstheworld.Withstraightforwardprotocolsandsimpleyethighlypredictiveendpoints,EpiOcularassaysareroutinelyutilizedthroughoutthecosmetic,personalcare,chemicalandpharmaceuticalindustries.Developedmorethan20yearsago,EpiOcularhasbeenusedfortestingthousandsofmaterials.Whetherscientistsneedariskassessmentassayfortheirritationpotentialofanoculardeliveredmaterialorthehazardassessmentofchemicalbeingtransportedacrosstheworld,EpiOcularisthetestsystemofchoicetomeetyouroculartestingneeds.

OcularIrritationTests

EpiOcularEyeIrritationTest

TheEpiOcularEITisaninvitrohazardassessmentassaythathastheABIlitytodifferentiatematerialsthatareocularnon-irritantsfrommaterialsthatarenotocularirritants.TheEITisapplicabletoabroadrangeoforganicandinorganicchemicals,andisacceptedasOECDTG492.

EpiOcularEyeIrritationTestProtocol

WatchavideobyJoveonEpiOcularEIT

EpiOcularSubDraizeUltra-MildnessTest

TheEpiOcularUltra-mildnessTestisaninvitroriskassessmentassayroutinelyutilizedformildmaterialsforwhichtheDraizetestisinsensitive.ThishighlyreproducIBLeassayallowsforquantifiablediscriminationamongmild,milderandmildestproductformulations.

EpiOcularSubDraizeProtocol

EpiOcularMTTET-50(Neat)

Thisinvitroriskassessmentassayisbasedontheeffectivetimeatwhichamaterial(appliedneat)causesa50%reductionintissueviability(ET-50).BasedontheET-50,thetestarticleiscategorizedintooneof4classificationsrangingfromnon-irritatingtosevere/extremewhichcorrespondtogroupingsofRabbitDraizeEyeScores(MMAS).

EpiOcularMTTET-50NeatProtocol

EpiOcularMTTET-50(Dilution)

SimilartotheMTTET-50Neatmethod,thisinvitroriskassessmentassayisbasedontheeffectivetimeatwhichamaterial(dilutedto20%inwater)causesa50%reductionintissueviability(ET-50).BasedontheET-50,thetestarticleiscategorizedintooneof4classificationsrangingfromnon-irritatingtosevere/extremewhichcorrespondtogroupingsofRabbitDraizeEyeScores(MMAS).Thismethodisrecommendedforsurfactantbasedsolutionsandisapplicabletowater-solublematerialswithaspecificgravityof>0.95.

EpiOcularMTTET-50DilutionProtocol

BrowseourreferencelibrarytoseehowourEpiOculartissuehasbeenusedintheseareasofstudy.

Tissue:

Kit:StandardEpiOcular™kit(OCL-200)consistsof24tissues,eachtissue9mmindiameter.(Tissue“kits”containtissues,asmallamountofculturemedium,andplasticware;contactMatTekforspecifickitcontents.)

Substrate:Non-coated,9mmIDsinglewelltissuecultureplateinsertsareused(e.g.MillicellPCF,Nuncpolycarbonatesinglewelltissuecultureplateinserts).

Culture:Atairliquidinterface.

Histology:5-8celllayers(basallayertoapicalsurface).

Cornification:None.

LotNumbers:Tissuelotsproducedbyeachtechnicianforeachweekareassignedaspecificlotnumber.Typically,therearemultiplelotnumbersforanygivenweek’stissueproduction.Aletterofthealphabetisappendedtotheendofthelotnumbertodifferentiatebetweenindividualkitswithinagivenlotoftissues.Alltissuekitswithinalotareidenticalinregardstocells,medium,handling,cultureconditions,etc.

Shipment:At4°Conmedium-supplemented,agarosegelsin24-wellplate.

Shipmentday:EveryMonday.ShipmentonThursdayalsopossibleuponspecialrequest.

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

Shelflife:Includingtimeintransit,tissuesmaybestoredat4°Cforupto4dayspriortouse.However,extendedstorageperiodsarenotrecommendedunlessabsolutelynecessary.Inaddition,thebestreproducibilitywillbeobtainediftissuesareusedconsistentlyonthesameday,e.g.Tuesdayafternoonorfollowingovernightstorageat4°C(Wednesdaymorning).

Alternativetissues:

OCL-200-ABF:Antibiotic-freetissue.Forlast3daysofculture,gentamicinisomittedfromculturemedium.Customerreceives200mlofOCL-200-ASY-AFABmedium.

OCL-200-AFF:Anti-fungal-freetissue.Forlast3daysofculture,Amphotericinisomittedfromculturemedium.Customerreceives200mlofOCL-200-ASY-AFABmedium.

OCL-200-AFAB:Antibiotic,anti-fungal-freetissue.Forlast3daysofculture,gentamicinandAmphotericinareomittedfromculturemedium.Customerreceives200mlofOCL-200-ASY-AFABmedium.

OCL-200-EACH:SingleEpiOculartissueinsert.

OCL-200-FRZN-EA:SingleOCL-200tissuethathasbeensubjectedto3freeze/thawcycles.DesignedtoserveascontrolforMTTauto-reductiontest.

OCL-212:Halfkit(12culturesinsteadofstandard24).

OCL-606:SameasOCL-200except6tissuesperkit,eachtissueculturedinalargercellcultureinsert(22mmdiameter).

Cells:

Type:Normalhumanepidermalkeratinocytes(NHEK).

Geneticmake-up:Singledonor.

Derivedfrom:Neonatal-foreskintissue.

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

Medium:

Basemedium:Dulbecco’sModifiedEagle’sMedium(DMEM).

Growthfactors/hormones:Epidermalgrowthfactor,insulin,hydrocortisoneandotherproprietarystimulatorsofnon-keratinizingepithelialdifferentiation.

Serum:None.

Antibiotics:Gentamicin5µg/ml(10%ofnormalgentamicinlevel).

Anti-fungalagent:AmphotericinB0.25µg/ml.

pHIndicator:Phenolred.

Alternatives:Phenolred-free,antibiotic-free,anti-fungal-free,orhydrocortisone-freemediumandtissueareavailable.Agentsareremovedatleast3dayspriortoshipment.

Maintenancemedium:MostassayswithEpiOcular™areperformedwithin24hours.However,forlongerexperiments,thetissuecanbemaintainedinOCL-200-MM(identicaltoEpiOcular™assaymedium).

QualityControlandSterility:

Visualinspection:Alltissuesarevisuallyinspectedandifphysicalimperfectionsarenoted,tissuesarerejectedforshipment.

End-usetesting:Tissuesareexposedto0.3%TritonX-100for5,20,and60minutes.Thetimeofexposurerequiredtoreducethetissueviability(ET-50)usingtheMTTassayisdetermined(SeeMatTekEpiOcularTMET-50protocol)foreachlotoftissue.ET-50’smustfallwithintherangeofthe1996EpiOcular™database:12.2-37.5minutes.ET-50’sincustomers’labsareoftenverysimilartoMatTekresults.

Sterility:Allmediausedthroughouttheproductionprocessischeckedforsterility.Maintenancemediumisincubatedwithandwithoutantibioticsfor1weekandcheckedforsterility.Theagarosegelfromthe24-wellplateusedforshippingisalsoincubatedfor1weekandcheckedforanysignofcontamination.

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

Notificationoflotfailure:IfatissuelotfailsourQCorsterilitytesting,thecustomerwillbenotifiedandthetissueswillbereplacedwithoutcharge.BecauseourQCandsterilitytestingisdonepost-shipment,notificationwillbemadeassoonaspossible(Undernormalcircumstances,ET-50failureswillbenotifiedbyWednesday5pm;sterilityfailureswillbenotifiedwithin8daysofshipment).