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1、ArchivesofVirologyArchivesofVirology85,301--304(1985)©bySpringer-Verlag1985Replieationo[RatCoronavirusinaRatCellLine,LBCBriefReportByN.HIBANOl,K.0~O,Y.SADA1,A.I~ovs,1,T.MUBAKAMII,andH.TAKAMAtCU21DepartmentofVeterinaryMicrobiology,IwateUniversity,Morioka
2、,and2DepartmentofBacteriology,IwateMedicalUniversity,Mo~oka,JapanWith3FiguresAcceptedJanuary16,1985SummaryRatcoronavirusreadilypropagatedandinducedmarkedcytopathiceffectinaratcellline,LBCcellculture,whichprovidedasensitive,practicalassaysystemforviralin
3、fectivityandneutralizingantibody,andasatis-factorysourceofthevirus.PA~KERetal.(4)haveisolatedanagentwithcharacteristicmorphologyofeoronavirus(5)fromthelungofrats.Thisratcolonywaspositiveforcomplementfixingantibodyagainstmousehepatitisvirus(MHV).Theagent
4、,designatedasrateoronavirus(RCV),wasshowntosharecommonantigen(s)withMHV.ByintranasMinoculationwithI~CV,afatalpneu-monitiswasproducedinnewbornrats.ThegrowthofRCVhasbeenreportedonaprimaryratkidneycellculturebutnotonanyestablishedcelllines.Becauseofthelack
5、ofthesusceptiblecelllinesforIgCV,consistentworkwithI~CVstillremaindifficult.Thisbriefcommunicationrecommendsaratcellline,LBC,providingpropagationofRCVandusefultoolforin-fectivityassay.RCV,strain8190,waskindlysuppliedbyDr.J.C.Parker,MicrobiologicalAssoci
6、ates,Bethesda,Maryland,U.S.A.TheLBCcelllinewasestablishedin1983byDr.K.Kai,InstituteofMedicalScience,UniversityofTokyo,fromaspontaneousmammarytumoroccurringinaLewisratbycon-302N.HI~ANo,K.O~o,Y.SADA,A.I~O~E,T.MURAKAMI,andH.TAKA=~ARU:ventionMmethods.Furthe
7、rdermisofthiscelllinearestillunderinvestiga-tion.TheLBCcellsweregrownat37°CinEagle'sminimumessentialmedium(MEM)containing10percentfetalcalfserum(FCS)andkanamyein(0.06mg/ml).TheFCSconcentrationwasreducedto5percentformaintainingthecellsorharvestingtheviru
8、s.Cellsgrownin50mlculturebottleswerewashedoncewithDulbeceo'sphosphatebufferedsalinepH7.2(PBS),andin-oculatedwith0.2mlofvirusmaterial.Aftervirusadsorptionat37°Cfor90minutes,theinoculatedculturesweregivenmaintenancemediumandin-cuba
