Abella N, Schelcher F, Delverdier M, Concordet D, Valarcher J F, Espinasse J, Cabanie P
Service d'Anatomie Pathologique, Ecole Nationale Vétérinaire, Toulouse, France.
Res Vet Sci. 1994 Sep;57(2):163-71. doi: 10.1016/0034-5288(94)90053-1.
Technical information to facilitate bovine blood treatment for optimum lymphocyte flow cytometry analysis is reported. Murine monoclonal antibodies CC8 and CC63 were used to identify phenotypes corresponding to bovine CD4 T cells and CD8 T cells. Blood samples collected in acid citrate dextrose (ACD) enhanced leucocyte subpopulation separation compared with ethylenediamine tetra-acetic acid, heparin and sodium citrate. To preserve bovine blood before immunophenotyping, samples collected in ACD may be kept at 22 degrees C or at 4 degrees C and should be analysed within 32 hours. For isolation of white blood cells, whole blood lysis was faster and gave the same results as Ficoll gradient separation 1.077 and Ficoll gradient separation 1.083. After immunophenotyping, blood could be stored at 4 degrees C if fixed with paraformaldehyde within seven days. Owing to diurnal variations, blood should be collected at a standard time of the day.
本文报道了有助于进行牛血处理以实现最佳淋巴细胞流式细胞术分析的技术信息。鼠单克隆抗体CC8和CC63用于识别与牛CD4 T细胞和CD8 T细胞相对应的表型。与乙二胺四乙酸、肝素和柠檬酸钠相比，采集于枸橼酸葡萄糖（ACD）中的血样增强了白细胞亚群的分离效果。为在免疫表型分析前保存牛血，采集于ACD中的样本可保存在22℃或4℃，且应在32小时内进行分析。对于白细胞的分离，全血裂解更快，且与密度梯度为1.077的Ficoll梯度分离法和密度梯度为1.083的Ficoll梯度分离法效果相同。免疫表型分析后，若在七天内用多聚甲醛固定，血液可保存在4℃。由于存在昼夜变化，血液应在一天中的标准时间采集。
