简介：ObjectiveTopresentanexperimentalmethodthatallowsisolationofgreaterepithelialridge(GER)andlesserepithelialridge(LER)cellsfrompostnatalratcochleaeusingacombinatorialapproachofenzymaticdigestionandmechanicalseparationandtoinvestigatearetrovirus-mediatedgenetransfertechniqueforitspossibleutilityinimmortalizationoftheGERandLERcelllines,inanefforttoestablishaninvitromodelsystemofhaircelldifferentiation.MethodsGERandLERcellsweredissectedfrompostnatalratcochleaeandimmortalizedbytransferringtheSV40largeTantigenusingaretrovirus.Theestablishedcelllineswereconfirmedthroughmor-phologyobservation,immunnocytochemicalstainingandRT-PCRanalysis.TheHath1genewastransferredintothecelllinesusingadenovirus-mediatedtechniquestoexploretheirpotentialtodifferentiateintohaircells.ResultsTheestablishedcelllineswerestablymaintainedformorethan20passagesanddisplayedmanyfeaturessimilartoprimaryGERandLERcells.Theygrewinpatchesandassumedapolygonalmorphology.ImmunostainingshowedlabelingbySV40largeTantigenandIslet1(aspecificmarkerforGERandLER).AllpassagesofthecelllinesexpressedSV40largeTantigenonRT-PCRanalysis.Thecellsalsoshowedthecapabilitytodifferentiateintohaircell-likecellswhenforcedtoexpressHath1.ConclusionRetrovirus-mediatedgenetransfercanbeusedinestablishingimmortalizedprogenitorhaircelllinesinnewbornrat,whichmayprovideaninvaluablesystemforstudyinghaircelldifferentiationandregenerationfornewtreatmentofsensoryhearinglosscausedbyhaircellloss.