PKH26标记的骨髓间质干细胞在阿尔茨海默病大鼠中的迁移  

Migration of PKH26-labeled mesenchymal stem cells in rats with Alzheimer′s disease

作　　者：李文玉[1] 金日龙[2] 胡兴越[1] 

机构地区：[1]浙江大学医学院邵逸夫医院神经内科,浙江杭州310016 [2]浙江大学医学院附属第一医院骨科,浙江杭州310003

出　　处：《浙江大学学报（医学版）》2012年第6期659-664,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金项目(Y2090184);浙江省中医药局青年基金(2006Y005)

摘　　要：目的:研究荧光染料PKH26标记的骨髓间质干细胞(bone-marrow derived mesenchymalstem cells,BM-MSC)在阿尔茨海默病(Alzheimer's disease,以下简称AD)大鼠中的迁移。方法:抽取正常人骨髓,体外分离培养BM-MSC;以PKH26标记第5代BM-MSC,采用流式细胞仪检测BM-MSC表面标记物,并在荧光显微镜下观察PKH26标记率;将PKH26标记的BM-MSC尾静脉注射到正常对照组和AD动物模型组,14天后在大鼠海马区,用荧光显微镜观察PKH26标记的BM-MSC细胞。利用Morris水迷宫实验比较AD模型组和BM-MSC移植组空间学习记忆能力。结果:流式细胞仪检测BM-MSC表面标记物CD73和CD105呈阳性,在体外BM-MSC的PKH26标记率达100%。Morris水迷宫实验比较BM-MSC移植组和AD动物组,BM-MSC移植组在第13、14天时空间学习记忆能力比AD动物组有显著改善。在移植BM-MSC的大鼠海马区可发现PKH26标记的BM-MSC阳性细胞,与DAPI吻合。PKH26阳性细胞在AD动物模型组明显多于正常对照组。结论:BM-MSC不仅能通过AD大鼠的血脑屏障,而且存活在AD大鼠的海马区,并有能够改善AD模型大鼠的学习障碍。Objective： To investigate the migration of fluorescent dye PKH26-labeled BM-MSC in the Alzheimer′s model rats.Methods： Normal human bone marrow extracted for isolation of BM-MSC was cultured in vitro.The 5th passaged BM-MSC was labeled with PKH26,and observed under a fluorescence microscope for PKH26 labeling efficiency,and using flow cytometry BM-MSC surface markers was checked.The PKH26 labeled BM-MSC injected into the tail vein of the normal control group and AD animal model group,14 days after finding the PKH26-labeled BM-MSC cells in the rat hippocampus using fluorescence microscopy.Using the Morris water maze experiment comparison of AD model and BM-MSC transplantation group of spatial learning and memory ability.Results： Flow cytometry showed BM-MSC surface markers CD73 and CD105 were positive.In vitro,PKH26-labeled rate of BM-MSC was 100%.The Morris water maze experiment comparison of BM-MSC transplantation group and AD group of animals,BM-MSC transplantation group at 13,14 days of spatial learning and memory ability than AD animal group had significantly improved.14 days after BM-MSCs in rat hippocampus could be found which were PKH26-positive,consistent with DAPI staining.PKH26-positive cells in animal models of AD were significantly more than those in the normal control group.Conclusion： BM-MSC in AD rats not only migrates through the blood-brain barrier,but also mainly survives in the hippocampus of AD rats,and it can improve AD rat model of learning disabilities.

关 键 词：骨髓间质干细胞 骨髓细胞 阿尔茨海默病 PKH26 

分 类 号：R749.16[医药卫生—神经病学与精神病学]