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MatTek/MelanoDerm™/melanoderm/1 Ea
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Idealfortheassessmentofskinlighteningagentsandformulations,MelanoDermisusedbycosmeticscientistsworldwide.ReducethetimeandcostsassociatedwithclinicaltestingbyscreeningcompoundsandformulationsinMatTek’sMelanoDerm3Dtissuemodel.Differentiated3Dco-cultureofkeratinocytesandmelanocytesMelanocytesfromdifferentethnicitiesareavailableQuantitativereadoutswithin14daysDeliveredready-to-useTechnology:TissueCultureInsertMatTek’sMelanoDermSystemconsistsofnormal,human-derivedepidermalkeratinocytes(NHEK)andmelanocytes(NHM)culturedtoformamultilayered,highlydifferentiatedmodelofthehumanepidermis.TheNHMwithinco-culturesundergomelanogenesisleADIngtotissuepigmentation.MelanoDermisproducedusingserumfreemediumwithoutartificialstimulatorsofmelanogenesissuchasTPAandIBMX.Theculturesaregrownontissuecultureinsertsattheair-liquidinterface,allowingfortopicalapplicationofskinlightenersorself-tanningagents.MelanoDermprovidesausefulinvitromeanstoevaluatecosmeticandpharmaceuticalagentsdesignedtomodulateskinpigmentation.MelanoDermexhibitsinvivo-likemorphologicalandultrastructuralcharacteristicswhichareuniformandhighlyreproducIBLe.NHMlocalizedinthebasalcelllayerofMelanoDermaredendriticandspontaneouslyproducemelaningranuleswhichprogressivelypopulatethelayersoftissue.Whenculturedforupto3weeks,MelanoDermtissuesbecomeincreasinglypigmentedwithretentionofnormalepithelialmorphology.TissuescontainingNHMderivedfromblackdonorsshowincreasedpigmentationversusthosecontainingCaucasian-derivedorAsian-derivedNHM;allthreetypesofculturesaredistinctlydarkerthanNHM-freetissues(EpiDermTM).Thetopicalapplicationofknowninhibitorsofmelanogenesissignificantlyreducemelaninproductionandmacroscopicdarkening.Conversely,NHMwithinthetissuewillrespondtoknownstimulantsofmelanogenesis,suchasα-melanocytestimulatinghormoneandβ-fibroblastgrowthfactor,toproducetissueswhichdarkenfasterthanuntreatedcontrols.MelanoDerm’sstraightforwardassayshasbeenincorporatedasanintegraltestingstrategyinlaboratoriesfornearly20years.Withdozensoftechnicalreferencesavailable,thisgrowingbodyofdatademonstratesthatMelanoDermprovidesaninexpensiveandeffectivemeansofassessingskinpigmentationwhileavoidingspeciesextrapolationandtheuseoflaboratoryanimals.Applications:MelanoDermprovidesaninexpensiveandeffectivemeansofassessingskinpigmentationwhileavoidingspeciesextrapolationandtheuseoflaboratoryanimals.Simpleprotocolsandtheevaluationofearlycellularendpointsallowscientiststoacquiredataindays,notweeksormonths.SkinLighteningAquantitativeassaytodeterminemelanincontentinthetissueisperformedusingtheSOLVABLE™melaninassay.Thisassaycanbeusedtotesttheskinlighteningefficacyoffinalcosmeticformulations.SkinLighteningApplicationnotePigmentationStudiesMelanoDermhasbeenusedtostudythephysiologicalmechanismsofskinpigmentation.Searchourlibraryoftechnicalreferencesformoreinformation.a[rel~="mtli_filesize89716kB"]:after {content:" (897.16 kB)"}TechSpecs:Tissue:Kit:StandardMelanoDerm™kit(MEL-300)consistsof24individualtissues,eachtissue9mmindiameter.(Tissue“kits”containtissues,asmallamountofculturemedium,andplasticware;contactMatTekforspecifickitcontents.)Substrate:Chemicallymodified,collagen-coated,9mmIDsinglewelltissuecultureplateinsertsareused(e.g.MillicellCMorNuncpolycarbonatesinglewelltissuecultureplateinserts).Culture:Atairliquidinterface.Seedingratio:NHM:NHEK=1/10.Ratioof1/30availableuponrequest.Histology:8-12celllayers(basal,spinous,andgranularlayers).StratumCorneum:10-15layers(basedonTEM).LotNumbers:Tissuelotsproducedbyeachtechnicianforeachweekareassignedaspecificlotnumber.Typically,therearemultiplelotnumbersforanygivenweek’stissueproduction.Aletterofthealphabetisappendedtotheendofthelotnumbertodifferentiatebetweenindividualkitswithinagivenlotoftissues.Alltissuekitswithinalotareidenticalinregardstocells,medium,handling,cultureconditions,etc.Shipment:At4°Conmedium-supplemented,agarosegelsin24-wellplate.Shipmentday:EveryMonday.ShipmentonThursdayalsoavailableuponspecialrequest.Delivery:TuesdaymorningviaFedExpriorityservice(US).OutsideUS:Tuesday-Wednesdaydependingonlocation.Shelflife:Includingtimeintransit,tissuesmaybestoredat4°Cforupto6dayspriortouse.However,extendedstorageperiodsarenotrecommendedunlessnecessary.Inaddition,thebestreproducibilitywillbeobtainediftissuesareusedconsistentlyonthesameday,e.g.Tuesdayafternoonorfollowingovernightstorageat4°C(Wednesdaymorning).Lengthofexperiments:Culturescanbecontinuedforatleast2weekswithgoodretentionofnormalepidermalmorphology.Culturesmustbefedeveryotherdaywith5.0mlofeitherNewMaintenanceMedium(NMM)orNMM-113.Cellcultureinsertsareplacedatopculturestands(MEL-STND;seephotobelow)orwashers(EPI-WSHR)in6-wellplatestoallowuseof5.0ml. Alternativetissues:MEL-300-EACH:SameasMEL-300exceptonly1tissue,not24tissues.MEL-300-B-FRZN-EA:MEL-300-Btissuethathasbeensubjectedto3freeze/thawcycles.Designedtoserveascontrolformelaninassay.MEL-301:Cultured3dayslessthanMEL-300,lessdifferentiatedthanMEL-300MEL-312(-A,-B,-C):IdenticaltoMEL-300(-A,-B,-C)except12tissuesperkitinsteadof24.MEL-606(-A,-B,-C):SameasMEL-300(-A,-B,-C)except6tissuesperkit,eachtissueculturedinalargerdiametercellcultureinsert(ID=22mm).MEL-300-FT(-A,-B,-C):Full-thicknesshumanskintissuemodelcomposedoffibroblast-containingcollagenmatrix(dermis);normalhumanepidermalkeratinocytesandnormalhumanmelanocytes(epidermis).24tissuesperkit.Cells:Types:a)Normalhumanepidermalkeratinocytes(NHEK);b)Normalhumanmelanocytes(NHM).Geneticmake-up:NHEKandNHMarefromindividual,butdifferentdonors.TissuecontainingmatchedNHEKandNHMareavailableuponspecialrequest.Ethnicorigin:MelanoDermcontainingNHMfromCaucasian(MEL-300-C),Black(MEL-300-B),andAsian(MEL-300-A)donorsareavailable.Derivedfrom:Neonatal-foreskintissue.Alternatives:AlimitednumberofstrainsofNHEK&NHMfromadultbreasttissue.Screenedfor:HIV,HepatitisB,andHepatitisC.Medium:Basemedium:Dulbecco’sModifiedEagle’sMedium(DMEM).Growthfactors/hormones:Epidermalgrowthfactor,insulin,hydrocortisone,andotherproprietarystimulatorsofepidermaldifferentiation.Serum:None.Antibiotics:Gentamicin5µg/ml(10%ofnormalgentamicinlevel).Anti-fungalagent:AmphotericinB0.25µg/ml.pHIndicator:Phenolred.Otheradditives:Lipidprecursorsusedtoenhanceepidermalbarrierformation(proprietary).Alternatives:Phenolred-free(MEL-300-B-PRF),antibiotic-free(MEL-300-B-ABF),anti-fungal-free(MEL-300-B-AFF),andhydrocortisone-freemediumandtissue(MEL-300-B-HCF)areavailable.Agentsareremovedatleast3dayspriortoshipment.Maintenancemedium:Fourmaintenancemediaareavailable:Forlongtermexperimentsinwhichpigmentationandhistologyareimportant,theuseofEPI-100-NMM,EPI-100-NMM-113,orEPI-100-LLMMisrecommended.EPI-100-NMM(NewMaintenanceMedium)containsKGFwhichpreserveshistology,butdoesnotcontainanyfactorsthatdirectlypromotepigmentation.EPI-100-NMM-113containsKGF,β-FGF,andα-MSHandpromotespigmentationaswellasretentionofhistology.Longlifemaintenancemedium,EPI-100-LLMM,containsα-MSHandβ-FGF,butnotKGF.TissueswilldarkenmostrapidlyinLLMM,lessrapidlyinNMM-113,andleastrapidlyinNMM.ThebestretentionofhistologywillbeobtainedwhenNMMorNMM-113isused,andtheculturesarefed5.0mlofmediaevery48hr.The4thmedium,EPI-100-MM,isabasalmediumandthatcanbeusedforshorttermtoxicologicaltesting.Itdoesnotcontainanyfactorsthatpromotepigmentation.EPI-100-MMshouldnotbeusedforexperimentsinwhichtissuehistologyisanimportantendpoint.QualityControlandSterility:Visualinspection:Alltissuesarevisuallyinspectedandifphysicalimperfectionsarenoted,tissuesarerejectedforshipment.Selecttissuesareexaminedmicroscopicallytoinsurethatmelanocytesarevisible(forMEL-300-BandMEL-300-Atissues)andthattheyareevenlydistributedthroughoutthetissue.End-usetesting:TissuesarecontinuedinEPI-100-LLMMtoinsurethatmacroscopicdarkeningoftissuesoccurs.Tissuesareexposedto1%TritonX-100for5hrs.TheviABIlityat5hrsmustexceed45%asdeterminedbytheMTTassay(SeeMatTekEpiDerm™ET-50protocol)foreachlotoftissue.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,%viabilityfailureswillbenotifiedbyWednesday5p.m.;sterilityfailureswillbenotifiedwithin8daysofshipment).

MatTek公司是麻省理工学院两个化学工程教授于1985成立的科技公司。1991年,该公司利用其核心的聚合物表面改性技术进入新兴的组织工程领域,目前已成长为的组织工程技术公司,致力于生产创新性的3D重构人体组织模型,该模型相较于传统2D细胞模型具有更可靠的实验结果。其生产的重构皮肤,眼部和呼吸道组织模型已广泛应用于化妆品,化工,医药和家用产品等行业的毒理学监控(OECD,欧盟的指导方针)及毒理学研究。


MatTek公司的基于人体细胞的3D皮肤模型EpiDerm1993年研发上市,立即获得巨大的成功。从此MatTek通过以下方式彻底改变了基于人体细胞的体外模型市场:(1)低成本(降低50-80%的成本);(2)通过发表质量控制标准(同时具备阳性和阴性对照)以确保可为行业及监管机构接受的高可重复性水平;(3)优化研究人员的实验结果。通过创造性使用非动物的体外测试,融合降低(直接/间接)成本的强大协同效应及显著提高的产品质量,MatTek极大的降低产品成本、提高实验室效率及高质量的高可重复性产品,减少实验动物的使用,具有无可超越的长期组织可重复性;3-D仿真结构;高度分化的组织模型;代谢和分裂活性;源于正常的人体细胞;培养、操作简便;无血清培养;可定量的客观检测;物美价廉的优点。因而使得MatTek人细胞来源的体外组织模型被包括美国、加拿大、欧洲、日本和东亚的众多跨国公司与研究机构采用。


目前MatTek公司的产品主要包括EpiDerm(皮肤模型)、MelanoDerm(黑色素模型)、EpiOcular(眼周表皮模型)、EpiAirway呼吸道上皮模型)、EpiDermFT (全厚的皮肤模型EpiVaginal(阴道上皮模型)、EpiOral(口腔上皮模型)、 EpiGingivalTM(牙龈上皮模型)、Psoriasis tissue(牛皮癣组织模型)及树突细胞。


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