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目的:观察嗅鞘细胞对胚胎脊髓后角神经元突起生长的影响。方法:分离孕15d胚胎大鼠脊髓,取纵向分开的后半制备成细胞悬液,培养于:(1)原代培养的嗅鞘细胞单细胞层上;(2)嗅鞘细胞培养上清中;(3)原代培养的成纤维细胞单细胞层上;(4)单纯培养。24h后终止培养,行抗神经丝-200免疫细胞化学染色。每组随机取15个神经元,在LeicaQ-570图像分析仪中测量各组神经元的平均总突起长度及平均单个最长突起长度,进行组间比较。结果:神经元平均总突起长度(1)～(4)组分别为408.62±126.91滋m、336.24±106.34滋m、199.37±76.14滋m和64.61±24.49滋m;平均单个最长突起长度(1)～(4)组分别为180.13±83.54滋m、141.22±53.68滋m、107.29±43.35滋m和23.04±5.30滋m。第(1)、(2)组神经元平均总突起长度和平均单个最长突起长度明显高于第(3)、(4)组(P<0.01),而(1)、(2)组之间没有明显差异(P>0.05)。结论:与嗅鞘细胞共培养能明显促进胚胎脊髓后角神经元突起的生长速度。
OBJECTIVE: To observe the effects of olfactory ensheathing cells on neurite outgrowth in embryonic spinal cord. Methods: Spinal cord was isolated from embryonic 15 day embryos and separated longitudinally from the last half into suspension. The cells were cultured in: (1) primary cultured olfactory ensheathing cell monolayers; (2) cultured in the supernatant of olfactory ensheathing cells ; (3) primary cultured fibroblast single cell layer; (4) simple culture. Cultures were terminated after 24 hours and anti-neurofilament-200 immunocytochemistry was performed. Fifteen neurons were randomly selected from each group. The average total neurite length and the average length of the longest protuberance neurons in each group were measured in a Leica Q-570 image analyzer and compared between groups. Results: The average length of the neurons (1) ~ (4) were 408.62 ± 126.91μm, 336.24 ± 106.34μm, 199.37 ± 76.14μm and 64.61 ± 24.49μm, respectively. The average single longest protuberance length ) ~ (4) group were 180.13 ± 83.54 m, 141.22 ± 53.68 m, 107.29 ± 43.35 m and 23.04 ± 5.30 m. The average total length of neurites and the average length of single longest neurons in group (1) and group (2) were significantly higher than those in group (3) and group (4) (P <0.01) There was no significant difference (P> 0.05). CONCLUSION: Co-culture with olfactory ensheathing cells can significantly promote the growth of neurons in the dorsal horn of embryonic spinal cord.