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金鱼视网膜外植体神经突生长的培养基要求及牛磺酸的营养作用。

Medium requirements for neuritic outgrowth from goldfish retinal explants and the trophic effect of taurine.

作者信息

Cubillos S, Fazzino F, Lima L

机构信息

Laboratorio de Neuroquímica, Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Apdo 21827, Caracas 1020-A, Venezuela.

出版信息

Int J Dev Neurosci. 2002 Dec;20(8):607-17. doi: 10.1016/s0736-5748(02)00105-3.

Abstract

The use of culture media of known composition are necessary for studying the role of trophic molecules. Since most of the in vitro research on regeneration of the optic nerve has been performed in the presence of fetal calf serum, the aim of this study was to obtain a medium in which the neuritic outgrowth from post-crush goldfish retinal explants could take place without adding fetal calf serum. After the lesion of the optic nerve (10 days), the retina of goldfish was dissected and explants were cultured for 5 and 10 days in the absence or in the presence of fetal calf serum, at which time the neuritic outgrowth was determined. Various concentrations and combinations of glucose, albumin, calcium, HEPES and taurine were used. The highest neuritic outgrowth was observed in the presence of fetal calf serum, in which condition the amino acid taurine increased length and density of neurites. Media supplemented with albumin, calcium or HEPES did not modify the outgrowth of neurites from the explants. However, glucose favored the neuritic outgrowth in a bell-shaped manner, although fibers were thinner than those observed in the presence of fetal calf serum. Taurine did not stimulate outgrowth of neurites from explants growing in a medium with optimal concentrations of glucose, indicating that elements of the fetal calf serum are determinant for the trophic effect of taurine. The present results contribute to further studies, such as those related to the effect of taurine and of trophic factors derived from the optic tectum, which would be performed in the presence of a medium free of fetal calf serum.

摘要

使用已知成分的培养基对于研究营养分子的作用是必要的。由于大多数关于视神经再生的体外研究都是在胎牛血清存在的情况下进行的,本研究的目的是获得一种培养基,在不添加胎牛血清的情况下,使挤压后金鱼视网膜外植体能够发生神经突生长。视神经损伤(10天)后,解剖金鱼视网膜,将外植体在不存在或存在胎牛血清的情况下培养5天和10天,此时测定神经突生长情况。使用了不同浓度和组合的葡萄糖、白蛋白、钙、HEPES和牛磺酸。在胎牛血清存在的情况下观察到最高的神经突生长,在这种情况下,氨基酸牛磺酸增加了神经突的长度和密度。添加白蛋白、钙或HEPES的培养基并没有改变外植体神经突的生长。然而,葡萄糖以钟形方式促进神经突生长,尽管纤维比在胎牛血清存在时观察到的更细。牛磺酸并没有刺激在含有最佳浓度葡萄糖的培养基中生长的外植体的神经突生长,这表明胎牛血清的成分对于牛磺酸的营养作用是决定性的。目前的结果有助于进一步的研究,例如那些与牛磺酸和来自视顶盖的营养因子的作用相关的研究,这些研究将在不含胎牛血清的培养基中进行。

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