【摘要】 目的比较Ficoll-Paque梯度离心法与免疫磁珠法分离T淋巴细胞的效果差异,筛选有效的、优异的分选小鼠脾原始T淋巴细胞方法。方法分别采用Ficoll-Paque梯度离心法及Miltenyi Biotec免疫磁珠进行小鼠淋巴细胞分离。血小板计数器统计单核淋巴细胞数,苔盼蓝染色判断细胞存活率,流式细胞术鉴定CD4+CD62+T淋巴细胞纯度,并进行统计学分析。结果免疫磁珠法CD4+CD62+T淋巴细胞纯度较高为(66.27±3.52)%,Ficoll-Paque梯度离心法细胞纯度为(15.27±4.16)%,两者比较差异有统计学意义(P<0.01);但Ficoll-Paque梯度离心法获取淋巴细胞数(1.47±0.35)×108较免疫磁珠法的(0.67±0.26)×108多,两者比较差异有统计学意义(t=8.199,P<0.01),且Ficoll-Paque梯度离心法细胞存活率(93.9±2.6)%较免疫磁珠法的(89.9±2.9)%高,两者比较差异亦有统计学意义(P<0.05)。结论免疫磁珠法适合分离小鼠脾原始T淋巴细胞并用于细胞纯度要求高的免疫学实验,但细胞得率需进一步优化。更多还原

【Abstract】 Objective To compare the effect of isolation of T-lymphocyte of mice by Ficoll-Paque gradient centrifugation and immunomagnetic bead method.Methods Ficoll-Paque gradient centrifugation and Miltenyi Biotec immune magnetic beads were used for isolation of T lymphocyte in mice.Cell number were calculated by platelet counter,cell survival rate was judged by trypan blue dyeing,CD4+CD62L+T lymphocyte purity was detected by flow cytometry.Results The purity of CD4+CD62L+T lymphocyte in the immune magnetic bead method group was higher(66.27±3.52%),and lower in Ficoll-Paque gradient centrifugation group(15.27 ±4.16%),showing significant difference(P<0.01),but lymphocyte count in Ficoll-Paque gradient centrifugation group(1.47±0.35×108)was more than immune magnetic bead goup(0.67±0.26×108)showing significant difference(t=8.199,P<0.001).In addition,the cell survival rate in Ficoll-Paque gradient centrifugation group(93.9 ±2.6%)was higher than immunomagnetic bead group(89.9 ±2.9%)also showing significant difference(P<0.05).Conclusions Immunomagnetic bead method is suitable for cell purity demanding immunological experiment on condition that further optimization of the cell yield.更多还原