Study on the condition for central nerve regeneration-grafting of ependymal cells and cell proliferation by neuregulin-

神经调节素促进中枢神经再生-室管膜细胞移植及细胞增殖条件的研究

基本信息

批准号：
10470005
负责人：
IDE Chizuka
金额：
$ 8万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470005/
关键词：
nerve regeneration choroid plexus ependymal cell graft spinal cord coculture dorsal root ganglion neuron axonal outgrowth

项目摘要

1 Grafting into spinal cordThe choroid plexus from the 4th ventricle of adult rats (Wistar) were minced into small fragments and grafted into the dorsal funiculus at the C2 level in adult rat spinal cord from the same strain. Electron microscopy and fluorescence histochemistry showed that ependymal cells of the grafted choroid plexus intimately interacted with growing axons, serving to support the massive growth of regenerating axons. HRP injection at the sciatic nerve showed that numerous HRP-labeled regenerating fibers from the fasciculus gracilis extended into the graft 7 days after grafting. These regenerating axons from the fasciculus gracilis were maintained for at least 10 months, with some axons elongating rostrally into the dorsal funiculus. Evoked potentials of long duration were recorded at a level ca. 5mm rostral to the lesion in the rats 8 to 10 months after grafting. These findings indicate that choroid plexus ependymal cells have the ability to facilitate axonal growth in vivo, suggesting that they may be a promising candidate as graft for the promotion of nerve regeneration in the spinal cord.2 Coculture experimentChoroid plexuses from the 4th ventricle of postnatal day-1 to -5 mice, were mechanically dissociated and plated in bronectin-coated culture dishes. Ependymal cells spread in monolayer with few endothelial cells at 3-week culture. Dorsal root ganglia (DRG) were cocultured on the choroid plexus ependymal cell monolayer. It was demonstrated that neurons extended many long neurites with elaborate branchings on the surface of S100 β-stained ependymal cells. In contrast, DRG neurons cultured on the astrocytes and laminin-coated plates had much fewer and shorter primary neurites with fewer branches than those cultured on the ependymal cells. These results suggest that choroid plexus ependymal cells have a great capacity to promote neurite outgrowth from DRG neurons in vitro.

1 脊髓移植将成年大鼠(Wistar)第四脑室的脉络丛切成小片，移植到同一品系成年大鼠脊髓C2水平的背索中，电镜和荧光组织化学显示室管膜。移植的脉络丛细胞与生长的轴突密切相互作用，有助于支持再生轴突的大量生长。在坐骨神经处注射 HRP 显示，移植后 7 天，来自股薄束的大量 HRP 标记的再生纤维延伸至移植物中，这些来自股薄束的再生轴突维持了至少 10 个月，其中一些轴突向头端伸长至背侧。在距大鼠病变约 5mm 的水平处记录了长持续时间的诱发电位。移植后8至10个月。这些发现表明脉络丛室管膜细胞具有促进体内轴突生长的能力，表明它们可能是促进脊髓神经再生的有希望的候选者。2共培养实验脉络丛。将出生后第 1 天至第 -5 天小鼠的第 4 脑室的细胞机械分离并铺在铺有布连蛋白的培养皿中。培养3周后，将背根神经节（DRG）单层与脉络丛室管膜细胞单层共培养，结果表明神经元在S100 β染色的室管膜细胞表面延伸出许多长神经突。相比之下，在星形胶质细胞和层粘连蛋白包被的板上培养的 DRG 神经元比在星形胶质细胞和层粘连蛋白包被的板上培养的 DRG 神经元具有更少、更短的初级神经突和更少的分支。这些结果表明脉络丛室管膜细胞在体外具有促进 DRG 神经元神经突生长的巨大能力。