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1、PROCEEDINGSOFTHESOCIETYFOREXPERIMtNTALBIOLOGYANDMEDICINE161,183-185(1979)EffectofCyclicNucleotideonCoronavirusReplication(40516)ARNOLDS.MONTOANDH.F.MAASSABDepartmentofEpidemology,UniversityofMichigan,AnnArbor,Michigan48109Althoughthehumancoronaviruseshaveinoculatedint
2、otubeculturesofBS-C-1cells.beenestablishedasmajorpathogensoftheTubeswereincubatedat33"for5daysandrespiratorytract,relativelylittleisknownthematerialwasharvestedbyfreezing,afteraboutcharacteristicsoftheiractivityinmanadditionofgelatin,toafinalconcentrationorthetotalnum
3、berofdifferentstrainsthatof0.5%.Followingsixserialpassages,theexist(1-4).Thislackofinformationisdirectlyviruswasconsideredtobeadaptedandanaresultofdifficultyencounteredinisolationinfectiouspoolmade.Thetiterofthispoolandpropagationoftheagents.Oneofthewas105.5TCID50/ml.
4、viruses,229E,istheonlycoronaviruswhichCellculturesused.WI-38aswellastheBS-wasoriginallyisolatedincellcultureandisC-1cellswereemployed.TheWI-38cellsserologicallydistinctfromallothers(5).Sub-werereceivedastubecultures(HEMLabo-sequenttothefirstdescriptionof229E,iso-rator
5、ies,Rockville,MD);theBS-C-1cellslationofrelatedstrainshasbeenreported,werereceivedasbottlecultures(FlowLabo-oftenusingcellsnotgenerallyavailable.Inratories,Rockville,MD).Thecellswerese-contrast,noneoftheremainingcoronavirusriallypropagatedinourlaboratoryandtubetypesha
6、severbeenisolatedincellculture.culturesmadeweekly.GrowthmediumwasAllwereidentifiedbyvirtueoftheirgrowthEagle'sMinimumEssentialMedium(MEM)intrachealorganculture,andonlyoneofthiswith10%fetalbovineserum.Priortouse,thegroup,OC(organculture)43hasbeenBS-C-1cellswerewashedth
7、reetimeswithadaptedtoothersystems,firsttosucklingHanks'BalancedSaltSolution(BSS)andfedmousebrainandthentocellculture(6-8).withserumfreeEagle'sMEM.WI-38cellsThisviruscanbeconsideredasaprototypewerefedwithEagle'sMEMcontainingeitheroftheotherorgancultureviruses,sincea2%f
8、etalbovineserumor0.1%bovineserumnumberofthemarerelatedserologicallytoitalbuminfractionV.Variousadditiveswereandone,OC38,isid
