草鱼头肾淋巴细胞体外转化培养影响因素的研究

STUDIES ON SOME ELEMENTS OF TRANSFORMATION OF LYMPHOCYTES FROM HEAD-KIDNEY OF GRASS CARP (CTENOPHARYNGODEN IDELLUS)

从草鱼 (Ctenopharyngodenidellus)头肾中分离淋巴细胞 ,利用DMEM培养液 ,采用MTT比色法 ,测定草鱼头肾淋巴细胞在体外受特异有丝分裂原植物凝集素 (PHA)刺激下发生的转化率 ,研究在细胞培养过程中不同因素对淋巴细胞转化率的影响 ,以期建立一种较可靠的鱼类免疫学研究技术 .实验表明 ,头肾淋巴细胞体外增殖受PHA浓度、小牛血清浓度及温度条件影响 .PHA的最佳质量浓度为 80mg/L ,小牛血清的最佳体积分数为 15 % ,最适温度范围是 2 8- 30℃ .实验结果还表明 。

Lymphocytes were separated from head-kidney of grass carp (Ctenopharyngoden idellus) and incubated in DMEM medium. Stimulation indexes of lymphocytes from the head-kidney stimulated by PHA were determined by MTT colorimetry. The influence of the concentration of PHA(5, 10, 20, 40, 80, 160, 320 mg/L), concentration of calf serum (CS) (2%, 4%, 8%, 10%, 15%, 20%, 25%) and temperature (14, 18, 25, 30, 35, 37℃) on the stimulation indexes were studied. The aim was to set up a method in investigation fish immunity. The results showed that the optimum concentration of PHA was 80mg/L, the optimum concentration of CS was 15% and the optimum temperature was 30℃. The result also showed that the MTT colorimetry was suitable for measuring the transformation of lymphocytes from head-kidney in grass carp.