MatTek/Psoriasis/psoriasis/1 Ea

MatTek’sdiseased3Dmodelofpsoriasisisproducedfromnormalhumanepidermalkeratinocytesandpsoriaticfibroblastsharvestedfrompsoriaticlesions.Thecellsareculturedonspeciallypreparedcellcultureinsertsusingserumfreemediumtoformamultilayered,highlydifferentiatedtissue.ThePsoriasistissuesmaintainapsoriaticphenotypeasevidencedbyincreasedbasalcellproliferation,expressionofpsoriasis-specificMarkersandelevatedreleaseofpsoriasis-specificcytokines.Morphologically,thetissuemodelcloselyparallelslesionalpsoriatichumantissues.Thismodelprovidesresearcherswithahumanrelevant,highlycontrolledinvitromodeltoassessthesafetyandefficacyofleadtherapeuticcompoundsandtostudyotherbasicpsoriasisBIOLOGyphenomena.

Thepsoriasistissuemodelexhibitsapsoriaticphenotypeasevidencedbyincreasedexpressionofpsoriasis-associatedmarkersincludinghumanβ-defensin-2(HBD2),psoriasin,SKALP/elafin,keratinocytehyperproliferativecellsandproinflammatorycytokines/chemokinessuchasIL-6,IL-8,GM-CSF,andIP-10. Additionally,MatTek’sPsoriasismodelcanbeproducedfrom6differentdonorsallowingforthestudyoftherapeuticswhileaccountingforbiologicalvariABIlitybetweendonors.

TheinvitroPsoriasismodelisresponsivetomarketedanti-psoriatictherapeuticsandisamenabletomodelskin–immunesysteminteractionswiththeexogenousadditionofsolubleimmunecellfactors.

Anti-PsoriasisDrugScreening

Themodelprovidesresearcherswithauseful,invitromeanstoassessthepre-clinicalsafetyandefficacyofleadtherapeuticcompounds.

PsoriasisApplicationNote

PsoriasisDiseaseResearch

Thetissuemodelcloselyparallelslesionalpsoriatichumantissuesintermsofmorphologyandcytokineexpression.Thismodelprovidesresearcherswithauseful,invitromeanstostudypsoriasisbiologyphenomena.

Searchourlibraryoftechnicalpapersformoreinformation.

Tissue:

Kit:ThePsoriasisTissueKit(SOR-300-FT)consistsof24tissues.

Substrate:MilliporeMillicell™Cellcultureinsertused.Poresize=0.4μm,Diameter=0.8cm,Surfacearea=0.6cm2.

Culture:Atairliquidinterface(apicaltissuesurfaceexposedtotheatmosphere).

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

LotNumbers:Tissuelotsareproducedeveryotherweekandareassignedaspecificlotnumber.Aletterofthealphabetisappendedtotheendofthelotnumbertodifferentiatebetweenindividualkitswithinagivenlotoftissues.Alltissuekitswithinalotareidenticalinregardstocells,medium,handling,cultureconditions,etc.

Shipment:At2-8°Conmedium-supplemented,agarosegelsin24-wellplates.

Shipmentday:EveryotherMonday.

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

Shelflife:Includingtimeintransit,tissuesmaybestoredintheiroriginalpackageat2-8°Cforupto4dayspriortouse.Thebestreproducibilitywillbeobtainediftissuesareusedconsistentlyonthesameday,e.g.Tuesdayafternoonorfollowingovernightstorageat2-8°C(Wednesdaymorning).Extendedstorageperiodsarenotrecommended.

Lengthofexperiments:Culturescanbecontinuedforupto2weekswithgoodretentionofnormalepidermalmorphology.Culturesmustbefedeveryotherdaywith5.0mlofSOR-300-FT-MMmedium.Culturestands(MatTekpart#:MEL-STND)arerequiredforexperimentsinexcessof24hrs.

Alternativetissues:

SOR-300-FT-ABF:Antibiotic-freetissue.Forlast3daysofculture,gentamicinisomittedfromculturemedium.CustomeralsoreceivesSOR-300-FT-MM-AFABmedium.

SOR-300-FT-AFF:Anti-fungal-freetissue.Forlast3daysofculture,Amphotericinisomittedfromculturemedium.CustomeralsoreceivesSOR-300-FT-MM-AFABmedium.

SOR-300-FT-AFAB:Antibiotic,anti-fungal-freetissue.Forlast3daysofculture,gentamicinandAmphotericinareomittedfromculturemedium.CustomeralsoreceivesSOR-300-FT-MM-AFABmedium.

SOR-300-FT-CTR:Normalhealthycontroltissueforpsoriasisexperiments

Cells:

Type:Normalhumanepidermalkeratinocytes(NHEK);psoriatichumandermalfibroblasts(PHDF).

Derivedfrom:Neonatal-foreskintissue(NHEK);adultpsoriaticexplants(PHDF)

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

Medium:

Basemedium:Dulbecco’sModifiedEagle’sMedium(DMEM).

Growthfactors/hormones:Epidermalgrowthfactor,insulin,hydrocortisoneandotherproprietarystimulatorsofepidermaldifferentiation.

Serum:None.

Antibiotics:Gentamicin5µg/ml(10%ofthenormalgentamicinlevel).

Anti-fungalagent:AmphotericinB0.25µg/ml.

pHIndicator:Phenolred.

Otheradditives:Lipidprecursorsusedtoenhanceepidermalbarrierformation(proprietary).

Alternatives:Phenolred-free(SOR-300-FT-PRF),antibiotic-free(SOR-300-FT-ABF),anti-fungal-free(SOR-300-FT-AFF),orhydrocortisone-freemediumandtissue(SOR-300-FT-HCF)areavailable.Agentsareremoved3dayspriortoshipment.

Assay/Maintenancemedium:SOR-300-FT-MMisusedforequilibrationoftissuesfollowingshipmentandformaintenanceoftheSOR-300-FTtissuesincultureduringexperiments.

QualityControlandSterility:

Visualinspection:Alltissuesarevisuallyinspectedandifphysicalimperfectionsarenoted,tissuesarerejectedforshipment.

End-usetesting:Tissuesareexposedto1%TritonX-100for4,6,8and10hours.Thetimeofexposurerequiredtoreducethetissueviability(ET-50)usingtheMTTviabilityassayisdetermined(SeeMatTekPsoriasisTissueuseprotocol)foreachlotoftissue.ET-50’sgenerallyfallwithintherangeof6.0-12.0hours.ET-50’sincustomers’labmaydifferslightlyfromtheMatTekresults.Inaddition,qPCRdataforpsoriasisrelatedgeneswillbemonitored.Increasesof>5foldforpsoriasin,humanbeta-defensin-2andelafinareexpected.

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-50failureswillbenotifiedbyThursday5pm;sterilityfailureswillbenotifiedwithin8daysofshipment).