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关于Shh协同骨髓间充质干细胞促进造血干细胞增殖机制的研究的开题报告.docx

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关于Shh协同骨髓间充质干细胞促进造血干细胞增殖机制的研究的开题报告 Title:MechanismsofShh-mediatedenhancementofhematopoieticprogenitorcellproliferationbyco-culturingwithbonemarrow-derivedmesenchymalstemcells Introduction: Hematopoieticstemcelltransplantation(HSCT)isacommonlyusedtherapyformanyhematologicaldisorders,suchasleukemia,lymphoma,andaplasticanemia.However,thelimitedavailabilityofcompatibledonorsandthecomplicationsassociatedwithtransplantprocedureshaveledtothesearchfornew,accessible,andeffectivetherapiestoenhancethehematopoieticfunction.Co-culturinghematopoieticstemcellswithbonemarrow-derivedmesenchymalstemcells(BMMSCs)hasbeenshowntoenhancetheproliferationanddifferentiationofhematopoieticstemcells.AmongthevarioussignalingpathwaysinvolvedinBMMSCs-mediatedhematopoieticstemcellmaintenanceandrenewal,SonicHedgehog(Shh)signalingpathwayplaysanessentialrole. Objectives: TheaimofthisstudyistoinvestigatethemechanismsofShh-mediatedenhancementofhematopoieticprogenitorcellproliferationbyco-culturingwithBMMSCs.Thespecificobjectivesare: 1.ToverifytheexpressionofShhligandanditsreceptorPtch-1inBMMSCsandhematopoieticprogenitorcells. 2.TodeterminetheeffectofShhsignalingpathwayactivationontheproliferationanddifferentiationofhematopoieticprogenitorcellsinco-culturewithBMMSCs. 3.ToelucidatethedownstreamsignalingpathwaysinvolvedinShh-mediatedenhancementofhematopoieticprogenitorcellproliferationbyco-culturingwithBMMSCs. Methods: 1.TheexpressionofShhligandanditsreceptorPtch-1willbeinvestigatedbyRT-PCRandimmunofluorescencetechniquesinBMMSCsandhematopoieticprogenitorcellsseparately,andintheco-cultures. 2.Hematopoieticprogenitorcellswillbeco-culturedwithBMMSCswithorwithoutadditionoftheShhagonist,SAG.Theproliferationanddifferentiationofhematopoieticprogenitorcellswillbeassessedbyflowcytometry,colony-formingunitassays,andgeneexpressionanalysisofhematopoieticstemcellmarkers. 3.ThedownstreamsignalingpathwaysinvolvedinShh-mediatedenhancementofhematopoieticprogenitorcellproliferationbyco-culturingwithBMMSCswillbeinvestigated