摘要
目的 探讨移植的胚胎小脑组织在成年小鼠脑内 ,黑质细胞和TH转基因细胞移植在猴脑内 ,人胚脑细胞植入人脑内能否存活及观察植入的神经元能否纠正和改善中枢神经系统的功能缺陷和症状。方法 (1)将胎龄 13d的C5 7BL/ 6小鼠胚胎小脑组织悬液植入 3个月龄的BALB/c成年小鼠小脑内。 (2 )将构建的TH基因修饰细胞及胚脑黑质细胞植入猴帕金森病模型脑尾状核内。术后进行组织学观察。在此基础上 ,将胎龄为 9～ 12周的人胚胎小脑细胞悬液植入到 6例重度小脑萎缩病人的小脑内 ,并观察疗效。选用自动流产人胚黑质细胞及肾上腺髓质细胞悬液移植到 5例晚期帕金森病病人脑尾状核内。结果 小鼠及猴的动物实验组织学证实神经细胞移植后可在受体脑内存活和再生 ,并有浦氏细胞分化成熟和迁徙到颗粒细胞层。TH基因修饰细胞移植到猴的尾状核内 ,可以纠正帕金森病猴的症状和体征 ;移植 6个月后组织切片表明猴帕金森病模型脑微包囊内转基因细胞存活而且TH基因表达呈强阳性。 6例重度小脑萎缩病人及 5例晚期帕金森病病人移植后近期疗效良好 ,多数病例远期疗效退步 ,仅有 2例病人疗效稳定。结论 未成熟的神经细胞或转基因细胞在受体脑内可以存活 ,并具有神经细胞再生和改善神经功能的潜力。
Objective To study the survival ability of cerebellar cells and genetically modified cells transplanted in the brains of mice and monkeys and whether the transplanted neurons correct and improve the cerebral function disorders. Methods Suspension of cerebellar tissue from 13 day old C57BL/6 mouse embryos was transplanted into the cerebellum of 3 month old BALB/c mice with Purkinji cell degeneration. Since one week after the transplantation, six mice were killed every week till the 8 th week to examine the cerebellar tissue by immunohistochemistry. Genetically modified neuroblastoma cells with tyrosine hydroxylase (TH) gene and substantia nigra cells of naturally aborted 12 week old human embryo were implanted into the caudatum nucleus of two monkeys with Parkinson′s disease caused by injection of 1 methy 4 pheny 1, 2,3,6 tetrahydropyridine. After the operation, the monkeys′ symptoms were observed. Six months later, the monkeys were killed. The transplanted areas were examined histologically and immunohistochemically. On the basis of the animal experiment, cerebellar tissue suspension of 9 to 12 week old human embryos was transplanted into the cerebellum of 6 patients with severe cerebral atrophy, and suspension of substantia nigra cells and adrenal medullary cells from naturally aborted human embryos was transplanted into the caudatum nucleus of 5 patients with late Parkinson′s disease. Then the patients were followed up to observe the symptoms. Results Surviving transplanted embryonic cerebellar cells could be seen in the cerebelli of recipient mice. Differentiation, maturation, and migration into the granular layer of Purkinje cells could be seen. Axons and dendrites grew from the newly generated Purkinje cells. Many mature nerve cells were monoclone antibody staining positive. The myodynamia of monkeys increased and tremor was alleviated, and torsion spasm was remarkably remitted 5 and 7 days after the transplantation respectively. Genetically modified cells with tyrosine hydroxylase gene could be seen and were pale brown with immunohistochemical staining in the cerebral microcapsule of monkeys. Five of the 6 patients with severe cerebellar atrophy and 5 patients with late Parkinson′s disease showed improvement of symptoms beginning from 2 weeks after transplantation. The average score of Webster′s scale in the patients with Parkinson′s disease decreased from 21 to 11. Th3 condition of two patients with cerebellar atrophy and 2 patients with Parkinson′s disease continued toimprove or remained stable after two years′ follow up. The condition of other patients had worsened. Conclusion The immature embryonic nerve cells and transgenic TH cells survive in the brain of recipients and have the potential to regenerate neurons and improve the function of central nervous system.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2002年第7期440-444,共5页
National Medical Journal of China
基金
国家自然科学基金资助项目 (3 9670 75 1)
