鸡T淋巴细胞和B淋巴细胞分离纯化方法的建立

将鸡胸腺和腔上囊淋巴组织剪碎后与PBS磷酸盐缓冲液混合,通过密度梯度离心和尼龙棉柱过滤对T、B淋巴细胞进行了分离纯化。纯化的T、B淋巴细胞以FITC标记的鼠抗鸡CD4单抗,RPE标记的鼠抗鸡CD8单抗,FITC标记的鼠抗鸡Bu-1单抗染色,用流式细胞仪分类检测其纯度,用台盼蓝排斥试验检测T、B淋巴细胞的活力。结果表明,以本研究建立的方法分离到的T、B淋巴细胞纯度较高(均达90%以上),细胞活力强,与流式细胞仪分选的效果相当,且经济简便。

Establishment of simple method for purification of T and B lymphocytes of chicken

Lymphoid tissues from thymus and bursa Fabricus of chicken were sheared and mixed with PBS buffer.T lymphocytes and B lymphocytes isolated from the lymphoid tissues were purified by density gradient centrifugation and nylon wool chromatography and stained with mouse-anti-chicken's CD4-FITC,CD8-RPE and Bu-1-FITC monoclonal antibodies,respectively.The purity and activity of the T and B lymphocytes were examined using flow cytometry and trypan blue exclusion assay,respectively.The results showed that the purity of T and B cells purified with the above mentioned method was more than 90% and their activities were similar to that of the cells isolated with flow cytometry.