欢迎访问《医学研究杂志》网站！

PKH26荧光示踪剂在神经干细胞移植大鼠创伤性脑损伤中的应用

投稿时间：2010-04-09 修订日期：2010-05-10 点此下载全文

引用本文：马海英,李花,孔力,喻博,石玉秀.PKH26荧光示踪剂在神经干细胞移植大鼠创伤性脑损伤中的应用[J].医学研究杂志,2010,39(7):39-41

DOI：

摘要点击次数: 1557

全文下载次数: 1191

作者	单位
马海英	中国医科大学组织胚胎学教研室、大连医科大学组织胚胎学教研室
李花	中国医科大学组织胚胎学教研室
孔力	大连医科大学组织胚胎学教研室
喻博	中国医科大学附属盛京医院
石玉秀	中国医科大学组织胚胎学教研室

基金项目:国家自然科学基金资助项目 (30850001)；辽宁省教育厅科学技术研究项目 (2008851,2008779)

中文摘要:目的探讨神经干细胞移植创伤性脑损伤(TBI)过程中有效的示踪剂，并进一步观察移植后NSCs的自然存活与分化情况。方法Feeney法制备大鼠TBI模型；将体外培养的NSCs进行PKH26荧光示踪剂标记，并于损伤后第3天移植入脑损伤灶区，对照组注射等量生理盐水。分别于移植后1天、4天、7天及2周、3周、4周、8周取材，行全脑冷冻切片，观察PKH26标记的NSC的自然存活及分布情况；Nestin、βⅢ-微管蛋白和GFAP免疫细胞化学染色观察移植后NSCs的分化。同时，于取材前采用Corner试验进行神经行为学检测。结果PKH26标记NSCs的阳性率＞95%。移植后第1天、4天，NSCs主要分别于注射部位周围，细胞胞体较小，分布均匀，大部分仍为Nestin阳性表达细胞。移植后第7天，移植NSCs可迁移至嗅球、额叶及枕叶皮质等区域。针道及周围组织NSCs分化为神经元和神经胶质细胞的比例分别为12.3％±2.1％和372％±7.6％。移植后第2周，海马、脑桥、小脑普肯耶细胞层等广泛区域均可见移植细胞。移植后第4～8周，移植细胞存活数量呈减少趋势。移植细胞的PKH26染色未见明显减弱。同一时间段内，与对照组相比，NSCs移植组动物的感觉运动功能均有明显改善。结论PKH26荧光示踪剂可用于体外培养的NSCs脑内移植示踪观察。移植NSC于损伤灶区，可自然分化为神经元和神经胶质细胞,并可迁移至脑内多处远距离部位，能明显改善TBI动物的感觉运动功能。

中文关键词:PKH26 神经干细胞移植创伤性脑损伤

Neural Stem Cells Labeled by Exposure to the Fluorescent Dye PKH26 Delivery to the Damaged Cortical Areas in the Traumatically Injured Brain of Rats

Abstract:ObjectiveTo determine if PKH26 could be used to serve as an effective tracer and observe the ability of transplanted neural stem cells to different into neural cells and astrocytes during traumatic brain injury. MethodsWe transplanted NSCs labeled by exposure to the fluorescent dye PKH26 into the cortical areas at 3 days after traumatically injured brain survey. The brains of rats were collected at 1, 4, 7days, 2, 3, 4 and 8 weeks after transplantation.The grafted cells were identified by their immunoreactivity to nestin for NSCs, βⅢ-tubulin for neuron and GFAP for astrocytes. We evaluated experimental neurology by corner test. ResultsThe results revealed the positive ratio of cells labeled by PKH26 was more than 95%. The majority of the surviving cells were located within the implantation sites at 1,4 days after transplantation. Most of these cells were nestin+. A small proportion of cells were βⅢ-tubulin+(12.3%+2.1%), and GFAP+ was detected in 37.2%±7.6％ of transplanted cells at 7days after transplantation. Many transplanted NSCs migrated extensively outside the transplantation sites, including hippocampus, pons and Purkinje cell layer of cerebellum at 2 weeks after transplantation. From 4 weeks to 8 weeks after transplantation, the surviving NSCs were decreased, but PKH26 was nondiffusion. Transplanted NSCs improved sensorimotor function after experimental traumatic brain injury. ConclusionPKH26 fluorescent tracer can be used to observe NSCs transplanted into the brain. Transplanted NSCs can survive in the traumatically injured brain and differentiate into neurons and astrocytes, migrate away from the graft core, and improve sensorimotor function after experimental traumatic brain injury.

keywords:PKH26 Neural stem cell Transplantation Traumatic brain injury (TBI)

查看全文查看/发表评论下载PDF阅读器