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1、Jou~alofVirolagicaimethod,25(1989)63-7063ElsevierJVM00896GrowthofamurinecoronavirusinamicrocarriercellculturesystemPierreJ.Talbotl,JacquesLapierre2,ClaudeDaniel,RobertDugrk2andPierreTrCpanier2Cenrrederechercheenvirologieand2Divisiondesvaccinsviraux,InstitutArmand-Fra
2、ppier,Universitt!duQutbec,VilledeLaval.QuCbec,Canada(Accepted23February1989)SummaryThegrowthofthemurinecoronavirusMHV-A59onmurineDBTcellsadaptedtodextran-madeCytodex1microcarrierswasstudiedincomparisonwithcellsgrownonplasticdishes.Withamicrocarrierconcentrationof5g/l
3、inspinnerflasks,adensityof3xlo6cells/mlwasreachedin7days.Undertheseconditions,cellssupportedvirusgrowthtothesameextentaswhentheyweregrownontheplasticsubstratum.Thiswasshownbyasimilardevelopmentofvirus-inducedsyncytia,thereleaseofanequivalentnumberofinfectiousprogenyv
4、irionspercell,similarrecoveriesobservedafterconcentrationandpurificationandanidenticalappear-anceofthepurifiedvirusundertheelectronmicroscope.Ontheotherhand,thetechnicalconvenienceofmicrocarriersandtheeaseofscale-upemphasizetheirpotentialforthegrowthofcoronaviruses.C
5、oronavirus;Microcarrier;CellcultureIntroductionCoronavirusesaretheetiologicalagentsofvariousacuteandchronicdiseasesofmammalsandfowl(Wegeetal.,1982;Siddelletal.,1983).Theyareenvelopedvirusesof80to120nmindiameterandcontainsingle-strandedRNAofpositivepolarity.Inmiceandr
6、ats,severalstrainsofmurinehepatitisviruses(MHV)areCorrespondenceto:Pierre3.Talbot,InstitutArmand-Frappier,UniversitCduQuCbec,531,bout.desPrairies,ViliedeLavai,Quebec,H7VlB7Canada.0~66~934/89/$03.5001989ElsevierSciencePublishersB.V.(BiomedicalDivision)responsibleforne
7、urological,respiratoryandgastrointestinaldisorders.WearestudyingtheneurotropicA59andJHMstrainsofMHV,asanimalmodelsofvi-rus-inducedneurologicdisease,withpossiblerelevancetohumandisorders.Mo-leculardeterminantsofpathogenesiswerelocatedontheviralspikeglycoproteinE2(Talb
8、otetal.,1988aandb).Tocharacterizefurtherthisprotein,theproteinwaspurifiedfromMHV-A59virionsbyaffinitychromatographyfromconcen-trate
