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1、MaterialsandMethodsMouseModelsAllanimalworkwascarriedoutunderproceduralandethicalguidelinesoftheBritishHomeOffice.TodeterminethecontributionoftheBMtohepaticstellatecellsandmyo-fibroblastsduringthedevelopmentofcirrhosis,weperformedsexmismatchedBMtransplantationsfrommaledonormiceintof
2、emalerecipients;6-week-oldfemaleBalb/cmicereceivedlethalirradiation(8Gyinadivideddose4hoursapart)andwholeBMtransplantsfrom6-week-oldmaledonors.MiceimmediatelyreceivedatailveininjectionofBM;unlessstated,thiswas1_106wholeBMcellsisolatedfromflushingthefemur,tibia,andpelvisofmaledonormi
3、cewitha29-gaugeneedlecontainingphosphate-bufferedsaline(PBS)/2%fetalcalfserum(FCS).Micewereplacedonacidifiedwater,and,4weekslater,micereceivedintraperitoneal(IP)injectionsof1_LpergrambodyweightofaCCl4/oliveoilmixture(1:7ratio,Sigma-Aldrich,Gillingham,UnitedKingdom)every5days.Groupso
4、fmice(n_4unlessstated)werekilledatintervalsfrom0to12weeksofCCl4,alwaysat72hoursfollowingthelastinjection.TodeterminewhetherBM-derivedstellatecellsandmyo-fibroblastswereastablecellpopulationaftertherecoveryofliverinjury,BMtransplantedmicethathadreceived8weeksofCCl4wereallowedtorecove
5、rfor8weekspriortotissueanalysis.Asecondmodelofliverdamagewasalsoused:femaleBalb/cmicereceivedmaleBMtransplantsasbefore;4weekslater,TAA(Sigma,T-8531)wasadministeredIPat200mg/kgbodyweight(dilutedindistilledwater)3timeseachweekfor4weeks.MicereceivingTAA(n_8)andcontrols(nodamage,n_4)wer
6、ekilled,andtissuewasharvested3daysafterthefinaldoseofTAA.CellsofBMoriginweretrackedinliversectionsthroughtheuseoffluorescentinsituhybridization(FISH)fortheYchromosome.Inaddition,toconfirmtheFISHanalysisintissue,maleandfemalecontrolmiceandanumberofmicethathadreceivedBMtransplantsand8
7、weeksofCCl4hadstellatecellsisolatedfromtheirlivers,usingcollagenaseandpronasedigestionfollowedbydensitycentrifugation.25FISHwasperformedontheisolatedstellatecells.ToassesswhethertheBM-derivedhepaticmyofibroblastswerecapableofintrahepaticcollagentranscription,6-week-oldfemaleC57/B6mi
8、ce(after10Gyirradia
