热休克胶质瘤源exosomes分离 鉴定 相关蛋白组成的研究及体外培养人树突状细胞-study on isolation and identification of related proteins from exo somes, a heat shock glioma source, and culture of human dendritic cells in vitro.docxVIP

徐州l医学院硕士学位论文热休克胶质瘤源exosomes分离、鉴定、相关蛋白组成的研究及体外培养人树突状细胞中文摘要..、目的培养胶质瘤细胞并对胶质瘤细胞进行热休克处理，分离纯化胶质瘤源也exosomes，进一步验证胶质瘤细胞可分泌exosomes研究热休克对胶质瘤细胞分峰的exosomes中相关蛋白的影响，为进一步研究胶质瘤免疫治疗及增强exosomes免疫活性及体外实验提供理论依据。通过体外培养人树突状细胞，为下…步构建DC疫茵奠定基础方法对胶质瘤U251细胞进行常规传代培养收集培养上消液，采用经典的差速离心法结合超滤离心纯化得到胶质瘤细胞分泌的exosomes和经热休克处理(42C，2小时)后胶质瘤细胞分泌exosomes(Heatshock创Exosom创且也Exo)，电镜观察两组exosomes形态。BCA法对两种侃osomes中总蛋白浓度进行测定，westerrn.blot鉴定相关蛋白分子HSP70，HSC70及细胞问粘附分子ICAM斗的表达。人外周血培养树突状细胞。结果胶质瘤细胞及经热休克处理后均可分泌exosomes，电镜形态光明显差别。BCA法检测蛋白浓度，经热休克处理细胞分泌的exosomes总蛋白浓度增加westem乍blot测定经热休克处理胶质瘤细胞分泌exosomes中HSP70，HSC70表达量增加，细胞|问粘附分子ICAM斗的表达无明显变化。结it:1.胶质瘤细胞可以分泌exosomes2.HS-Exo中HSP70，HSC70表达比exosomes更丰富，HS-Exo比exosomes具有更强的免疫活性3.体外成功培养人树突状细胞关键词胶质瘤exosomes热休克蛋白免疫泊疗Heatshockgliomassourceexosomesseparatedandidentifiedandrelatedproteinsconsistingofstudyandculturedhumandendriticcellinvitro..Abstracth4ObjectiveTocultivategliomascellsandgliomascellstoheatshocktreatment，separationandpurification，furtherverificationsourcegliomasexosomesgliom邸cellscansecrete，discussesexosomesheatshockofgliomascellssecretetheexosomesyieldandrelatedproteins，theinfluenceofgliomasimmunotherapyforfurtherresearchandm由anceexosomesimmunityactivityandinvitrostudiesprovidetheoreticalbasisoAndsuccessinvitropeopledendriticcells，forthenextst叩∞nstructDCvaccinelaythefoundation.MethodU251cellsforgliomaroutinenestintrainingcollecttrainingsup臼natantfluid，usingclassicaldifferentialcentrifugationcombiningultrafiltrationpurifiedbygliomascel1ssecretetheexosomesandtheHeatshocktreatment(42.C，2hours)afterexosomes(cellssecretegliomasHeatexosomesshocked，HS-Exo)，electronmicroscopetwogroupsexosomesform.TwoexosomesBCAmethodforquantitativedetection，westerrn-blotappraisalrelatedproteinmoleculesHSP70.HSC70andbetweenthecelladhesionmoleculesICAM-lexpressionResultGlialcellsandtheheatshocktreatmentafterallcanse口eteexosomes，therewasnoevidentdifference.BCAmethodtodetectproteinconcentration，theheatshocktreatmentcellssecreteproteinsaresignificantlyincreased.Bytestingthethermalshockwesterrn-bl