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  • 简介:Thestudyofneuronalplasticityunderpathologicalconditionsisnowamajorpointoffocusonthefieldofneurologicalrecovery.Aftertherepeatedfailureofacuteneuroprotectionstrategiesforstroketreatment,thedesignofstudiesaimedatpromotingthereconstructionofneuronalnetworkshasbecomeessential.Methodsforthedeliveryoftherapeuticagentsonasteadydosage,thuspreventingpharmacologicalpeaksorexcessivemanipulationofexperimentalanimals,arethusrequired.Additionally,methodsthatallowthevisualizationofneurologicalremodelingprocessesarefundamentaltotheunderstandingofhowatherapeuticagentexertsitsfunction.Herewedescribehowtheuseofminiosmoticpumpsforthesteadydeliveryofsuchagents,togetherwithtracttracerinjections,canbecombinedtounveilimportantinformationonhowthebrainchangesafterstrokeandhowtherapeuticagentspromotebrainremodelingrecovery.

  • 标签: 神经元 可塑性 连通性分析 神经康复 大脑 治疗药物
  • 简介:BACKGROUND:Withdevelopmentsoftissueengineeringandgeneticengineering,weaimtoculturemyoblasts,whicharecharacterizedbyhighpurity,highqualityandhighproduction,forwideapplicationinneuralregenerationresearches.OBJECTIVE:Tomodifytraditionaldissociationmethodinordertoobtainmyoblasts,whicharecharacterizedbyhighpurity,highqualityandhighproduction,andexplorethebiologicalpropertiesunderinvitroculture.DESIGN:Observationalstudy.SETTING:BasicInstituteofAcademyofMilitaryMedicalSciencesofChinesePLA.MATERIALS:FourneonatalWistarratsof5daysold,bothgendersandmeanbodymassof10gwereselectedinthisstudy.Themainreagentsanddevicesweredetailedasfollows:DMEMmedium(GibcoCompany),fetusbovineserum(FBS,HycolneCompany),collagenaseⅡ(SigmaCompany),trypsin(SigmaCompany),dispaseⅡ(SigmaCompany),desminantibody(FuzhouMaixinCompany),antibodyⅡandABCkit(WuhanBosterBiotechnologyCompany),deskcentrifuge(KUBATO,Japan),andinvertedphasecontrastmicroscope(LEICADMIRB,Germany).METHODS:TheexperimentwascarriedoutintheBasicInstituteofAcademyofMilitaryMedicalSciencesofChinesePLAfromJunetoOctober2006.Neonatalratsweresacrificedundersterileconditiontoobtainskeletalmusclesoflimbs,whichwerewashedwithcoldPBS(containingbenzylpenicillinandestreptomicina),andmusculartissuewasshearedintopieces.Then,thosemuscularpieceswereaddedwithmixeddigestiveenzyme(containing2g/LcollagenaseⅡ+5g/LdispaseⅡ+0.28g/LCaCl2)astwicevolumeaspieces,dealtwithmechanicalpipettingfor5minutesandculturedinCO2incubatorfor10minutes.Theoperationwasdoneforthreetimesandthemuscularpiecesweredigestedfor45minutesintotal.Moreover,cellsweresuspendedagaininordertoobtainmyoblastsfromskeletalmuscleofneonatalrats.Inaddition,myoblastswerepurifiedwithdifferentialattachmenttechniqueandenzymedigestionsoastoobservemorpho

  • 标签: 大鼠 成肌细胞培养 改良方法 混合酶消化法 双纯化法