动物外周血单个核细胞分离方法探讨

目的 探索用人淋巴细胞分离液分离多种属动物外用血单个核细胞（PBMCs）的可行性及评价其分离效果,建立一种适于野外大规模多种属动物PBMCs的分离方法.方法 采用人淋巴细胞分离液分离来源于新疆的伊犁马、新疆驴、不同品种犬、新疆双峰驼、天山马鹿和来源于重庆的荣昌猪、中国荷斯坦奶牛、简阳大耳黑山羊外周血PBMCs.随机抽样检测分离的动物PBMCs细胞总数、细胞纯度和细胞活率.结果 用人淋巴细胞分离液成功分离上述8种动物PBMCs,每毫升动物外周血分离的PBMCs细胞总数为0.52×106～2.03×106个,纯度为67%～93%,细胞活率为92.5%-98.0%.结论 用人淋巴细胞分离液分离多种动物PB-MCs的方法宜在动物的病毒分子流行学调查中进一步推广.

Discussion on Method for Separation of Animal Peripheral Blood Mononuclear Cells

Objective To establish a available method to separate the peripheral blood mononuclear cells （PBMCs） from many kinds of animals adopting human lymphocyte separation medium. Methods The separation of PBMCs in the blood of Ill horses, Xinjiang donkeys, canines,Xinjiang Bactrian Camels and Tianshan red deer from Xinjiang and Rongchang pigs, Chinese Holstein cow, Jianyang Big-ear Black goats from Chongqing city is performed by means of human lymphocyte separation medium. The total number, cell purity and cell survival rate of PBMCs are tested at random. Results The total number of PBMCs isolated from the above-mentioned eight kinds of animals is 0.52 × 10^6 ～2.03× 10^6/mL peripheral blood;cell purity of PBMCs is 67% -93% ;cell survival rate is 92.5% - 98.0%. Conclusion The method above,which shows significant priority, moreover,can be widely used in the molecular epidemiology investigation of virus in animals.