冠状病毒Coronavirusl论文-1983 Clones of MRC-C cells may be superior to the parent line for the culture of 229E-like strains of human respiratory.pdf

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1、JournalofVirologicalMerhods,6(1983)267-269261ElsevierCLONESOFMRC-CCELLSMAYBESUPERIORTOTHEPARENTLINEFORTHECULTUREOF229E-LIKESTRAINSOFHUMANRESPIRATORYCORONAVIRUSR.J.PHILLPOTTSCommorrColdUnit.CoombeRoad,Salisbury,Wilts.U.K.(Accepted18February1983)Aclonewasselectedfromthe

2、MRC-CcontinuousheteroploidcelllinewhichwassignificantlybetterthanMRC-Cforthecultureofhumanrespiratorycoronavirus(HCV)229E.strainLP.SuchclonescouldprovegenerallyusefulfortheisolationofHCV-229Efromclinicalspecimens,andforthepropagationandassayoflaboratoryadaptedstrains.

3、cloningcelllinehumanrespiratorycoronavirusMRC-Cisacontinuousheteroploidcelllinederivedfromhumanembryolungwhichhasbeenusedfortheisolationofhumanrespiratorycoronavirus229Estrains(HCV-229E)attheCommonColdUnit(Larsonetal.,1980).Itmaintainspoorlyinrollertubecultures,andoft

4、enlosesitssensitivitytoHCV-229Eathighpassagelevels.Heteroploidcelllinesmayhavemarkedlyunstablecharacteristics(Schneblietal.,1977)andcloneswithdifferingsensitivitiestoagivenvirusmaybeisolatedfromasinglehigh-passagecellline(seeforexampleClarkeandSpier,1980).Ittherefores

5、eemedlikelythatacloneofMRC-Ccellscouldbeselected,whichwouldprovesuperiortotheparentlinefortheisolation,propagationandassayofHCV-229E.AnumberofcloneswerederivedfromMRC-Cbythecultureofsinglecellsinmicrotitreplatewells,inHamsF12mediumsupplementedwith10%foetalcalfserum.Th

6、erewasconsiderablevariationinthegrossmorphologyoftheresultingculturesandappearanceswhichvariedfromfibroblastic(approximately75%ofcultures)toepithelioid,weremaintainedonsubculture.Clonesrepresentativeofallmorphologicaltypes,andMRC-Cwerecomparedinparallelexperimentsfors

7、ensitivitytoHCV-229EstrainLPbothbyquanta1assayofanasalwashpooltakenfromvolunteerschallengewithLP,andplaqueassayofapoolofMRC-CadaptedLP.TheresultsoftheseexperimentsareshowninTable1.Quanta1assaytitresrangedfrom

8、ellcellsmaintainedinrollertubeculture.However,goodmaintenancedidnotappeartobeassociatedwithsensitivitytoHCV-229Estrain0166-0