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借鉴哺乳动物内皮细胞体外培养技术 ,结合鱼类细胞自身特点 ,纯化培养鲤血管内皮细胞并传至 3代 ,以此作为粘附材料。应用淋巴细胞分离液离心技术并结合玻璃粘附法分离纯化鲫、草鱼外周血中淋巴细胞、单核细胞 ,并与上述 3代内皮细胞进行免疫粘附试验 ,同时进行免疫粘附动力学观察。结果显示 ,鲫、草鱼两类细胞对内皮细胞的粘附率分别为 :0 .0 9± 0 .0 13,0 .2 0± 0 .0 18;0 .11± 0 .0 15,0 .2 1± 0 .0 2 3。表明鲤科鱼类不同种的同类细胞粘附率差异不大 ,而同种不同类细胞则差异显著。动力学观察分析表明 ,淋巴细胞粘附较快 ,60min进入平台期 ;单核细胞粘附慢 ,12 0min进入平台期。初步证明鱼类内皮细胞具有免疫介导作用。
Draw lessons from mammalian endothelial cells in vitro culture technology, combined with the characteristics of fish cells, purified cultured carp vascular endothelial cells and passed to 3 generations, as an adhesive material. Lymphocytes and monocytes were isolated and purified from peripheral blood of crucian carp and grass carp by lymphocyte separation liquid centrifugation combined with glass adhesion method. Immunoadherence assay was performed with the above three generations of endothelial cells, and immunoadsorption kinetics was also observed. The results showed that the adhesion rate of Carassius auratus and grass carp cells to endothelial cells were: 0.09 ± 0.013, 2.020 ± 0.018, 0.101 ± 0.0. 2 1 ± 0 .0 2 3. The results showed that there was no significant difference in the adhesion rate of cytoplasm among different species of cyprinid species, while the difference of the same kind of cells was significant. Kinetic analysis showed that lymphocytes adhered rapidly and reached plateau at 60 min. Monocytes adhered slowly and reached plateau at 120 min. Preliminary evidence that fish endothelial cells have immune mediation.