简介：Parthenogeneticembryonicstemcellshavepluripotentdifferentiationpotentials,akintofertilizedembryo-derivedembryonicstemcells.Theaimofthisstudywastocomparetheneuronaldifferentiationpotentialofparthenogeneticandfertilizedembryo-derivedembryonicstemcells.Beforedifferentiation,karyotypeanalysiswasperformed,withnormalkaryotypesdetectedinbothparthenogeneticandfertilizedembryo-derivedembryonicstemcells.SexchromosomeswereidentifiedasXX.Immunocytochemistryandquantitativereal-timePCRdetectedhighexpressionofthepluripotentgene,Oct4,atboththemRNAandproteinlevels,indicatingpluripotentdifferentiationpotentialofthetwoembryonicstemcellsubtypes.Embryonicstemcellswereinducedwithretinoicacidtoformembryoidbodies,andthendispersedintosinglecells.SinglecellsweredifferentiatedinN2differentiationmediumfor9days.Immunocytochemistryshowedparthenogeneticandfertilizedembryo-derivedembryonicstemcellsbothexpresstheneuronalcellmarkersnestin,βIII-tubulinandmyelinbasicprotein.Quantitativereal-timePCRfoundexpressionofneurogenesisrelatedgenes(Sox-1,Nestin,GABA,Pax6,Zic5andPitx1)inbothtypesofembryonicstemcells,andOct4expressionwassignificantlydecreased.NestinandPax6expressioninparthenogeneticembryonicstemcellswassignificantlyhigherthanthatinfertilizedembryo-derivedembryonicstemcells.Thus,ourexperimentalfindingsindicatethatparthenogeneticembryonicstemcellshavestrongerneuronaldifferentiationpotentialthanfertilizedembryo-derivedembryonicstemcells.

简介：Ciliaryneurotrophicfactoristheonlyknownneurotrophicfactorthatcanpromotedifferentiationofhippocampalneuralprogenitorcellstoglialcellsandneuronsinadultrats.Thisprocessissimilartospontaneousdifferentiation.Therefore,ciliaryneurotrophicfactormaybeinvolvedinspontaneousdifferentiationofneuralstemcells.Toverifythishypothesis,thepresentstudyisolatedneuralprogenitorcellsfromadultmaleratsandculturedtheminvitro.Resultsshowedthatwhenneuralprogenitorcellswereculturedintheabsenceofmitogenfibroblastgrowthfactor-2orepidermalgrowthfactor,theyunderwentspontaneousdifferentiationintoneuronsandglialcells.Westernblotandimmunocytochemicalstainingshowedthatexogenousciliaryneurotrophicfactorstronglyinducedadulthippocampalprogenitorcellstodifferentiateintoneuronsandglialcells.Moreover,passage4adulthippocampalprogenitorcellsexpressedhighlevelsofendogenousciliaryneurotrophicfactor,andaneutralizingantibodyagainstciliaryneurotrophicfactorpreventedthespontaneousneuronalandglialdifferentiationofadulthippocampalprogenitorcells.Theseresultssuggestthatthespontaneousdifferentiationofadulthippocampalprogenitorcellsismediatedpartiallybyendogenousciliaryneurotrophicfactor.

简介：cAMPsignalingandthecontrolofSchwanncellfate:Theubiquitoussecondmessengercyclicadenosinemonophosphate(cAMP)controlsavarietyofcellularresponsesinacelltype-specificandstimulus-dependentmannerthroughanelaboratenetworkofsignalingintermediariesthatconnectstimulationofcellmembranereceptors(typicallyG

简介：BACKGROUND:Microgliaareverysensitivetoenvironmentalchanges,oftenbecomingactivatedbypathologicalconditions.Activatedmicrogliacanexertadualroleininjuryandrepairinvariousdiseasesofthecentralnervoussystem,includingcerebralischemia,Parkinson’sdisease,andAlzheimer’sdisease.OBJECTIVE:Animmortalmicroglialcellline,BV2,wastreatedwithvaryingconcentrationsoflipopolysaccharide(LPS)toinduceapathologicalsituation.Supernatantwasharvestedandincubatedwithbonemarrowmesenchymalstemcellsand,concomitantly,bonemarrowmesenchymalstemcelldifferentiationwasobserved.DESIGN:Acontrolledobservation,invitroexperiment.SETTING:DepartmentofNeurology,FirstAffiliatedHospitalofChinaMedicalUniversity.MATERIALS:Fivemale2–3-week-oldSpragueDawleyratswerepurchasedfromAnimalLaboratoryCenterofChinaMedicalUniversityandincludedinthisstudy.Theprotocolwasperformedinaccordancewithethicalguidelinesfortheuseandcareofanimals.ThemicroglialcelllineBV2wasproducedbyCellResearchInstituteofChineseAcademyofSciences.LPSwasproducedbySigmaCompany,USA.METHODS:ThisstudywasperformedintheCentralLaboratoryofChinaMedicalUniversityfromSeptember2006toMarch2007.Ratfemoralandtibialbonemarrowwascollectedforseparationandprimarycultureofbonemarrowmesenchymalstemcells.Bonemarrowmesenchymalstemcellculturesweredividedinto5groups:controlgroup,non-activatedgroup,aswellaslow-,medium-,andhigh-doseLPSgroups.Inthecontrolgroup,bonemarrowmesenchymalstemcellswereculturedwithDulbecco’smodifiedEagle’smedium(DMEM)supplementedwithfetalbovineserum(volumefraction0.1).Inthenon-activatedgroup,bonemarrowmesenchymalstemcellswereincubatedwithnon-activatedBV2supernatant.Inthelow-,medium-,andhigh-doseLPSgroups,bonemarrowmesenchymalstemcellswereincubatedwithLPS(0.01,0.1and1μg/L,respectively)-activatedBV2supernatant.MAIN

简介：BACKGROUND:ThedetectionofdifferentialgeneexpressioninbrainispossiblebycDNAmicroarraytechnology,andthescreeningofdifferentiallyexpressedgenesmightprovideabiologicalbasisforgene-targetedtherapyfortumors.OBJECTIVE:TodetectthedifferentialexpressionofgenesamongastrocytomaSHG-44(WHOgradeIV),CHG-5(WHOgradeII),andATRA-treatedSHG-44celllinesbycDNAmicroarray.DESIGN:Laboratoryexperimentsinvitro.SETTING:DepartmentofNeurobiology,theThirdMilitaryMedicalUniversity.MATERIALS:TheexperimentwasperformedattheDepartmentofNeurobiologyintheThirdMilitaryMedicalUniversityoftheChinesePLAfromJanuarytoOctober2007.TheSHG-44cellline(WHOgradeⅣ)wasestablishedbyProf.ZiweiDu,andtheCHG-5cellline(WHOgradeII)wassetupbyProf.XiuwuBianfromtheThirdMilitaryMedicalUniversityoftheChinesePLA.ThecDNAmicroarraycontaining9182knowngeneswaspreparedandprovidedbyDr.YangZhongattheCityUniversityofHongKong.METHODS:ToscreendifferentiallyexpressedgenesfromthegeneexpressionprofilesdetectedbycDNAmicroarraycomparisonsweremadebetweenCHG-5andSHG-44cellsandbetweenSHG-44cellswithorwithouttreatmentwith10μmol/LATRA.SomedifferentiallyexpressedgeneswereselectedrandomlyforNorthernBlotanalysistoconfirmtheresultsofthemicroarray.Thedeterminationcriteriafordifferentialgeneexpressionwereasfollows.①TheratioofCy5signaltoCy3wasgreaterthan2.0orlessthan0.5.②Theresultsofthetriplicatemicroarrayhybridizationsshowedthesametrendinthreeexperiments.③Ageneappearedatleasttwotimesonthetriplicatemicroarrayhybridizations,andthe3rdvaluedidnotshowacontradictorytrend.AnormalizedratioofCy5intensitytoCy3greaterthan2.0orlessthan0.5wasconsideredtorepresentup-regulatedordown-regulatedgeneexpression,respectively.MAINOUTCOMEMEASURES:Theidentificationofgenesthatweresimilarlyregulated(overlapping)dur

简介：Recentadvancesinstemcelltechnologieshaveopenednewavenuesforthetreatmentofanumberofdiseasesstilllackingeffectivetherapeuticoptions.Celltransplantationhasemergedasamongthemostpromisingclinicalinterventionfordisorderssuchasinjuries,diabetes,liver

简介：BACKGROUND:Culturesfrommultipleportionsofumbilicalcordbloodmesenchymalstemcellshavebeenshowntoundergomorerapidproliferationandattachmentthansingleportions.OBJECTIVE:Toobservegrowthofbasicfibroblastgrowthfactor(bFGF)-inducedculturesofhumanamnion-derivedmesenchymalstemcells(AMSCs)anddifferentiationintoneuronal-likecells.DESIGN,TIMEANDSETTING:Comparativeobservation.ThestudywasperformedattheLaboratoryofMicrobiologyandImmunology,BasicMedicalSchoolofZhengzhouUniversityfromJanuarytoMay2008.METHODS:Amniafromfull-term,uterine-incisiondeliveryweredonatedby12healthywomen.AMSCswereobtainedbycellseparationandculturetechniques,andwerepassagedandinducedbybFGF.Fromthethirdpassage,atotalof1mLAMSCs,atadensityof1.0×10~4/mL,wasseparatelyharvestedfromsixsamples,whichservedasgroupA.Atotalof1mLAMSCs,atadensityof1.0×10~4/mL,washarvestedseparatelyfromtheremainingsixsamples,whichservedasgroupB.Atotalof0.5mLfromthesixsamplesofgroupAand0.5mLfromthesixsamplesofgroupBwerecombinedtoformgroupC.MAINOUTCOMEMEASURES:Differencesincellquantityamongthethreegroupswerecomparedbycellquantificationand3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT)analysis.Expressionofaglialcellmarker,neuron-specificenolase,andnestinwasdetectedinthethreegroupsbyimmunocytochemistry.RESULTS:CellquantificationandMTTanalysisoflivecells,aswellasAMSCabsorbance,weresignificantlygreateringroupCcomparedwithgroupsAandBat18daysofculture(P<0.05),andnosignificantdifferencewasobservedbetweengroupsAandB.Glialfibrillaryacidicprotein,neuron-specificenolase,andnestinwereexpressedinallgroupsfollowingbFGFinduction.CONCLUSION:MixedAMSCculturespromotedproliferation,andbFGF-inducedAMSCsdifferentiatedintoneuronal-likecells.

简介：OurpreliminarystudiesconfirmedthatanactiveprincipleregionofBuyangHuanwudecoction,comprisingalkaloid,polysaccharide,aglycon,glucosideandvolatileoil,caninducebonemarrowmesenchymalstemcelldifferentiationintoneurons.Mitogen-activatedproteinkinasesignalingwasidentifiedasoneofthekeypathwaysunderlyingthisdifferentiationprocess.Thepresentstudyshowsphosphorylatedextracellularsignal-regulatedproteinkinaseandphosphorylatedp38proteinexpressionwasincreasedafterdifferentiation.Cellularsignalingpathwayblockingagents,PD98059andSB203580,inhibitedextracellularsignal-regulatedproteinkinaseandp38inmitogen-activatedproteinkinasesignalingpathwaysrespectively.mRNAandproteinexpressionoftheneuronalmarker,neuronspecificenolase,andneuralstemcellmarker,nestin,weredecreasedinbonemarrowmesenchymalstemcellsaftertreatmentwiththeactiveprincipleregionofBuyangHuanwudecoction.Experimentalfindingsindicatethat,extracellularsignal-regulatedproteinkinaseandp38inmitogen-activatedproteinkinasesignalingpathwaysparticipateinbonemarrowmesenchymalstemcelldifferentiationintoneuron-likecells,inducedbytheactiveprincipleregionofBuyangHuanwudecoction.

简介：BACKGROUND:Ithasbeendemonstratedthattransforminggrowthfactor-β(TGF-β)andbrain-derivedneurotrophicfactor(BDNF)caninducestemcelldifferentiationintoneuron-likecells.OBJECTIVE:ToinvestigatetheefficacyofTGF-βandBDNFatinducingthedifferentiationofadultratbonemarrowstromalcells(BMSCs)intoneuron-likecells,bothincombinationoralone.DESIGN,TIMEANDSETTING:AcomparativeobservationexperimentwasperformedattheDepartmentofOrthopedics,FirstAffiliatedHospitalofLiaoningMedicalUniversitybetweenOctober2007andJanuary2008.MATERIALS:TGF-βandBDNFwerepurchasedfromSigma,USA;mouseanti-ratneuronspecificenolase,neurofilamentandglialfibrillaryacidicproteinwerepurchasedfromBeijingHMHLBiochemLtd.,China.METHODS:BMSCswereisolatedfromratsaged4weeksandincubatedwithTGF-β(1μg/L)and/orBDNF(50μg/mL).MAINOUTCOMEMEASURES:Expressionofneuron-specificenolase,neurofilamentandglialfibrillaryacidicproteinweredeterminedbyimmunocytochemistry.RESULTS:BMSCsdifferentiatedintoneuron-likecellsfollowinginductionofTGF-βandBDNF,andexpressedbothneuron-specificenolaseandneurofilament.ThepercentofpositivecellswassignificantlygreaterinthecombinationgroupthanthoseinducedwithTGF-βorBDNFalone(P<0.01).CONCLUSION:TreatmentofBMSCswithacombinationofTGF-βandBDNFinduceddifferentiationintoneuron-likecells,withtheinductionbeingsignificantlygreaterthanwithTGF-βorBDNFalone.

简介：BACKGROUND:Stereotacticinjection(striatumorlateralventricle)andvascularinjection(tailveinorcarotidartery)arenowoftenusedincellulartherapyforcerebralinfarction.Stereotacticinjectioncanaccuratelydelivercellstotheinfarctarea,butrequiresastereotacticdeviceandcausessecondarytrauma;vascularinjectioniseasyandbetterforhostneurologicaldeficitrecovery,butcancausethrombosis.OBJECTIVE:Tocomparethetherapeuticpotentialofadultbonemarrow-derivedmesenchymalstemcells(BMSCs)transplantationbyintraperitonealversusintravenousadministrationtocerebralischemicrats.DESIGN,TIMEANDSETTING:ArandomizedcontrolledanimalexperimentwasperformedattheCellRoomandPathologyLaboratory,BrainHospitalAffiliatedtoNanjingMedicalUniversityfromNovember2007toSeptember2008.MATERIALS:BMSCswerederivedfrom20healthySprague-Dawleyratsaged4-6weeks.METHODS:Forty-fiveadultmiddlecerebralarteryocclusion(MCAO)ratswererandomlydividedintocontrol,intravenousandintraperitonealinjectiongroups,with15ratsineachgroup.At21daysaftermodeling,ratsinthecontrolgroupreceived1mLof0.01mol/Lphosphatebufferedsalineviatailveininjectionandeachexperimentalratreceived4×106BMSCslabeledbybromodeoxyuridine(BrdU)viaintravenousorintraperitonealinjection.MAINOUTCOMEMEASURES:Angiogeninexpressionandsurvivaloftransplantedcellsweremeasuredbyimmunohistochemicalstainingofbraintissueininfarctionhemisphereat7,14or21daysafterBMSCtransplantation.Co-expressionofBrdU/microtubule-associatedprotein2orBrdU/glialfibrillaryacidicproteinwasobservedbydouble-labeledimmunofluorescenceofcerebralcortex.Evaluationofnervefunctionusingtheneurologicalinjuryseverityscoreandtheadhesion-removaltestwasperformedonthe1stand21stdaybeforeandafterMCAO,andat3,7,14or21daysafterBMSCstreatment.RESULTS:Angiogenin-positivenewvesselsweredistributedinthebilateralstriatum,hippocampusandcerebral