中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9176-.doi: 10.3969/j.issn.1673-8225.2010.49.013

• 干细胞移植 • 上一篇    下一篇

经脑室植入神经干细胞在脑室周围白质软化新生大鼠脑内的迁移分化

贺月秋，陈惠金，钱龙华，陈冠仪   

  1. 上海交通大学医学院附属新华医院，上海市儿科医学研究所，上海市   200092
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 通讯作者: 陈惠金，博士生导师，主任医师，上海交通大学医学院附属新华医院，上海市儿科医学研究所，上海市 200092 hjchenk@online.sh.cn
  • 作者简介:贺月秋☆，女，1974年生，四川省泸定县人，汉族，2007年上海交通大学医学院毕业，博士，主治医师，现在成都市妇幼保健院新生儿科工作，主要从事早产儿脑损伤研究。 Eileenhe@163.com
  • 基金资助:

    上海市教委课题(05BZ08)。

Migration and differentiation of neural stem cells implanted into the ventricle in neonatal rats with periventricular leukomalacia

He Yue-qiu, Chen Hui-jin, Qian Long-hua, Chen Guan-yi   

  1. Shanghai Institute for Pediatric Research, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai  200092, China
  • Online:2010-12-03 Published:2010-12-03
  • Contact: Chen Hui-jin, Doctoral supervisor, Chief physician, Shanghai Institute for Pediatric Research, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China hjchenk@online.sh.cn
  • About author:He Yue-qiu☆, Doctor, Attending physician, Shanghai Institute for Pediatric Research, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China Eileenhe@163.com
  • Supported by:

    the Grant of Education Committee of Shanghai City, No. 05BZ08*

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摘要：

背景：对新生动物进行脑内神经干细胞移植，所植入神经干细胞的增殖、迁移、分化、轴突形成以及髓鞘化等能力，均远远高于成年动物。
目的：对经脑室植入神经干细胞在脑室周围白质软化新生大鼠脑内的迁移分化功能进行观察，探讨神经干细胞移植对治疗早产儿脑室周围白质软化的可行性。
方法：2日龄脑室周围白质软化新生大鼠在建模后72 h进行经脑室神经干细胞移植。外源性神经干细胞用PKH26荧光素标记。脑立体定位仪下经脑室穿刺部位：前-后=1.5 mm，中线－外侧=-2 mm，深度=1.5 mm；移植细胞浓度为5×1010 L-1；移植总量为2 μL；移植速度0.5 μL/min。应用激光共聚焦显微镜分别对植入神经干细胞于植入后1，2，3，7，14，21 d进行动态观察。并分别进行各分化细胞的免疫荧光分析。
结果与结论：应用激光共聚焦显微镜对植入神经干细胞进行动态观察，证实经脑室植入的外源性神经干细胞在脑内具有良好的迁移能力，3 d内大部分移行至脑室周围区域，并分布在损伤严重部位。2周左右，神经干细胞在脑室周围区域主要分化为OL前体，部分分化为神经元及星形胶质细胞。提示经脑室神经干细胞移植对早产儿脑室周围白质软化具有很大的治疗潜力。

关键词: 脑室周围白质软化, 少突胶质细胞前体, 神经干细胞, 细胞移植, PKH26

Abstract:

BACKGROUND: The abilities of proliferation, migration, differentiation, axon formation and myelinization of transplanted neural stem cells (NSCs) were greater in the brain of new-born animals compared with adult animals.
OBJECTIVE: To observe the migration and differentiation of exogenous NSCs transplanted into ventricle in neonatal rats with periventricular leukomalacia (PVL), and to explore the feasibility of NSCs transplantation for the treatment of PVL.
METHODS: The intraventricular transplantation of exdogenous NSCs labeled with PKH26 fluorescein was undertaken in two-day-old neonatal rats with PVL at 72 hours after operation. The ventricular puncture point under stereotaxis instrument was: AP=1.5 mm, ML=-2 mm, DV=1.5 mm. The cell density implanted was 5×1010/L; the volume dose was 2 μL and the velocity was 0.5 μL/min. NSCs implanted was dynamically observed under laser scanning confocal microscope and immunefluorescence analysis was undertaken for the confirmation of differential cells originated from NSCs at 1, 2, 3, 7, 14 and 21 days after transplantation, respectively.
RESULTS AND CONCLUSION: Laser confocal microscopy was used to observe transplanted NSCs, and confirmed that exogenous NSCs implanted through ventricle hold a favorable ability of migration. The most exogenous NSCs migrated into periventricular area and distributed in the serious injured region of white matter within three days after transplantation. Since two weeks after transplantation, NSCs implanted had differentiated mainly into OLs precursors, and partly into neurons or astrocytes in the periventricular area. These suggested that intraventricular transplantation of NSCs is with a great potential feasibility for the treatment of PVL.

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