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华西医学  2011 

骨髓间充质干细胞对大鼠到小鼠胰岛移植的保护作用

, PP. 641-645

Keywords: 骨髓间充质干细胞,异种胰岛移植,免疫调节

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Abstract:

【】 目的 探讨同种异基因骨髓间充质干细胞(bonemesenchamalstemcells,BMSC)静脉输注对大鼠到小鼠胰岛移植物的功能保护和小鼠糖尿病状态改善。 方法 全骨髓培养法获得C57BL/6小鼠BMSC。不连续梯度离心法分离纯化Sprague-Dawley(SD)大鼠胰岛,将300胰岛当量的胰岛单独或与BMSC联合移植入链脲菌素诱导的糖尿病BALB/c小鼠肾包膜下,并通过尾静脉在移植后0、3和5d注射CM-DiI标记的BMSC5×105/只,对照组给于磷酸盐缓冲溶液。移植后监测血糖,第9天处死小鼠,取肝、脾、胸腺、淋巴结和移植胰岛的肾脏,冰冻切片,荧光显微镜观察CM-DiI标记细胞的组织分布;免疫荧光法观察移植物中胰岛素和胰高血糖素表达,评价胰岛的功能。 结果 BMSC静脉输注后主要分布于胸腺,其次是脾脏和淋巴结,肾和肝组织中未观察到BMSC;BMSC联合胰岛移植组血糖控制水平优于其他组,且在第7天的口服糖耐量实验优于单纯胰岛移植组。 结论 与胰岛联合移植的BMSC对受者免疫器官和组织有明显的趋向性,且对胰岛细胞的体内存活有一定保护作用。【Abstract】 Objective Toresearchontheprotectionfunctionbytheallogeneicratbonemesenchymalstemcells(BMSC)onrattomouseislettransplantationandtheimprovementofdiabeticstateinmouse. Methods BMSCwerepreparedfromC57BL/6mousebonemarrowcellsandidentifiedbyflowcytometry(FCM).IsletswereisolatedfromSprague-Dawley(SD)ratswithFicolldiscontinuouscentrifugation.CM-DiIlabeledBMSCat5×105foronemousewereintravenouslyinfusedintoSTZinduceddiabeticBALB/cmiceafterrattomouseislettransplantationatday0,3and5.MicewithPBSintravenouslyinfusedafterislettransplantationweresetasthenegativecontrols.Bloodglucosewasmonitoredeverydayatthefirst3daysaftertransplantation,andthenmonitoredeverytwodays.Atday9aftertransplantation,spleen,thymus,lymphnods,liverandisletsrecipientkidneywereharvested.IcesliceswerepreparedandCM-DiIlabeledcellswereinvestigatedwithfluorescencemicroscope. Results CM-DiI-labeledBMSCweremainlydistributedinthymusfollowedbyspleenandlymphnodes.Inliverandkidney,therewasnoredfluorescenceobserved.ThebloodsugarcontrolforcombinedBMSCinfusiongroupwassuperiortoothergroups,andthecontrollevelofisletcombinedBMSCinfusiongroupwerebetterthansingleislettransplantationgroupinOGTTatday7. Conclusion AllogeneicBMSCcansustaintheinsulinsecretionofisletsinvivoandtendtodistributeinimmuneorgansoradenoidtissuesafterinfusion.

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