斑点叉尾[鱼回]头肾巨噬细胞分离及初步应用

Separation and Preliminary Application of Head Kidney Macrophages in Channel Catfish (Ietalurus punetaus)

通过Percoll密度梯度离心法分离斑点叉尾[鱼回]头肾巨噬细胞,经含2%胎牛血清的L-15培养液中培养,在光镜和电镜下观察其形态和超微结构;采用MTS法研究酵母细胞壁对巨噬细胞体外作用的合适浓度和最佳时间。结果表明:使用34%、51%的密度梯度分离斑点叉尾[鱼回]头肾细胞,经过3~12小时培养,巨噬细胞纯化率95%以上;分离的头肾巨噬细胞具有巨噬细胞的形态特征。酵母细胞壁对斑点叉尾[鱼回]的巨噬细胞最适的作用浓度为0-500pg/ml,作用时间为12小时。本研究为进一步探讨酵母细胞壁对鱼类免疫细胞的作用机理奠定了基础。

In this study, the head kidney macrophages of the channel catfish (Ietalurus punetaus) were isolated by Percoll continuous density gradient centrifugation and cultivated in L-15 medium with 2% fetal bovine serum.The morphological and ultrastructural features were observed under light and electron microscope. The appropriate concentration and optimal time of yeast cell wall on macrophages in vitro by MTS method. The results showed that the head kidney macrophages of channel catfish were isolated using a Percoll density gradient of 34% and 51%, more than 95% cells adhered and spread out on cover slips after 3?12 h in culture were macrophages.The obtained head kidney macrophages showed the morphological characteristics of macrophages. The appropriate concentration of the yeast cell wall on the head kidney macrophages were 0-500μg/ml, and the optimal time were 12 h. This work laid a foundation for further exploring the mechanism of yeast cell wall on fish immune cells.