Adult neural stem and progenitor cells modified to secrete GDNF can protect, migrate and integrate after intracerebral transplantation in rats with transient forebrain ischemia
Adult neural stem and progenitor cells modified to secrete GDNF can protect, migrate and integrate after intracerebral transplantation in rats with transient forebrain ischemia
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DOI：
10.1111/j.1460-9568.2007.05776.x
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发表时间：
2007-09-01
影响因子：
3.4
通讯作者：
Date, I.
中科院分区：
文献类型：
--
作者：
Kameda, M.;Shingo, T.;Date, I.
成人神经干细胞和祖细胞（NSPCs）可以分泌保护缺血脑的因子，是再生医学中重要的自体移植工具。我们研究了经基因修饰的成年NSPCs分泌更多的胶质细胞系来源的神经营养因子（GDNF）是否能保护大鼠的短暂性缺血。取成年Wistar大鼠脑室下区NSPCs，在表皮生长因子的作用下培养3周。用含有GDNF基因（NSPC-GDNF）或增强型绿色荧光蛋白（EGFP）基因（NSPC-EGFP；对照组）的纤维突变型Arg-Gly-Asp腺病毒处理NSPCs。在一个实验中，将培养的细胞移植到Wistar大鼠脑右侧缺血边界区。一周后，动物接受90分钟的腔内右大脑中动脉闭塞，随后进行磁共振成像和行为测试。与对照组相比，NSPC-GDNF组在术后1、7和28天的行为评分较高，梗死体积较小。在第二个实验中，我们在缺血损伤后3小时移植NSPCs。与对照组相比，移植后7天接受NSPC-GDNF的大鼠梗死面积减少，行为评估更好。第7天处死动物，取脑组织进行GDNF酶联免疫吸附测定和形态学评估。与对照组相比，更多的GDNF分泌，更多的NSPC-GDNF细胞向缺血核心迁移，更多的NSPC-GDNF细胞表达未成熟神经元标志物。此外，NSPC-GDNF组对小胶质细胞侵袭和凋亡有更有效的抑制作用。这些发现提示NSPC-GDNF可能在脑缺血治疗中有用。
Adult neural stem and progenitor cells (NSPCs) are important autologous transplantation tools in regenerative medicine, as they can secrete factors that protect the ischemic brain. We investigated whether adult NSPCs genetically modified to secrete more glial cell line-derived neurotrophic factor (GDNF) could protect against transient ischemia in rats. NSPCs were harvested from the subventricular zone of adult Wistar rats and cultured for 3 weeks in the presence of epidermal growth factor. The NSPCs were treated with fibre-mutant Arg-Gly-Asp adenovirus containing the GDNF gene (NSPC-GDNF) or enhanced green fluorescent protein (EGFP) gene (NSPC-EGFP; control group). In one experiment, cultured cells were transplanted into the right ischemic boundary zone of Wistar rat brains. One week later, animals underwent 90 min of intraluminal right middle cerebral artery occlusion followed by magnetic resonance imaging and behavioural tests. The NSPC-GDNF group had higher behavioural scores and lesser infarct volume than did controls at 1, 7 and 28 days postocclusion. In the second experiment, we transplanted NSPCs 3 h after ischemic insult. Compared to controls, rats receiving NSPC-GDNF had decreased infarct volume and better behavioural assessments at 7 days post-transplant. Animals were killed on day 7 and brains were collected for GDNF ELISA and morphological assessment. Compared to controls, more GDNF was secreted, more NSPC-GDNF cells migrated toward the ischemic core and more NSPC-GDNF cells expressed immature neuronal marker. Moreover, the NSPC-GDNF group showed more effective inhibition of microglial invasion and apoptosis. These findings suggest that NSPC-GDNF may be useful in treatment of cerebral ischemia.