CultureofsubconfluenthumanfibroblastsandkeratinocytesusingbiodegradabletransfermembranesC.Johnena,*,I.Steffena,D.Beichelta,K.Bra¨utigama,T.Witaschecka,N.Tomand,V.Moserd,C.Ottomannb,B.Hartmannb,J.C.Gerlacha,caCharite´,CampusVirchow-Clinic,DepartmentofSurgery,Universita¨tsmedizinBerlin,GermanybBurnCentre/DepartmentofPlasticSurgery,Unfallkrankenhaus,Berlin,GermanycDepartmentsofSurgeryandBioengineering,McGowanInstituteforRegenerativeMedicine,UniversityofPittsburgh,PA,USAdSchlossparkklinikBerlin,Germany1.IntroductionCell-freeandcell-containingskinsubstituteshavebeeninvestigatedanddevelopedforthetreatmentofmassiveburnsforseveraldecades.Althoughmanyskinsubstitutesarecommerciallyavailable,thatwhichprovidesthesamewidevarietyoffunctionsashumandermisandepidermishasnotyetbeendeveloped.Culturedkeratinocytegraftingisestablishedasatreatmentoptionforsevereburnsandchronicvenousulcers[1,2].Thecellsaregrownasmultilayersandtransferredassheets.Culturedepidermalautografts(CEAs)presentanumberofdisadvantages,includinglonginvitroexpansiontimes;3or4weeksarerequiredtoobtainsufficientcellstocovertheburnedsurface[3,4].Duringthistimethosewithmassiveburnsaresusceptibletosepsis,themajorcauseofmortalityamongthosewhosurvivetheinitialinjury[5].Thelengthofdelayinacutetreatmentisinverselyproportionaltothesizeofthebiopsyandtheefficiencyofcellexpansionintheculture,andisdirectlyproportionaltotheamountofmaterialrequiredforwoundcoverage[6].Inaddition,thedonor’sspecificmedicalhistorycanheavilyinfluencetheproliferationandgrowthofcellsintheculture[7].Furtherdisadvantagesofkeratinocytesheetslieintheirmechanicalfragilityandalsointheoccurrenceofhyperkeratosis,contractureandscarring[8].TheharvestingofCEAsrequiresenzymaticseparationfromthetissueculturesubstrate,whichmayresultincontractionandsubsequentrestrictedproliferation[9].Allogeneickeratinocyteshavebeenburns34(2008)655–663articleinfoArticlehistory:Accepted28August2007Keywords:SkinKeratinocytesFibroblastsExtracellularmatrixproteinsBiodegradableabstractT...
