流式细胞术检测不同试验条件对猪T 淋巴细胞亚群的影响

通过不同方式处理猪外周血淋巴细胞,对不同保存时间的淋巴细胞进行流式细胞术检测,探讨其对T淋巴细胞亚群中CD3^＋、CD4^＋、CD8^＋亚群的影响。采用肝素钠、EDTA、枸橼酸钠3种抗凝剂抗凝猪外周静脉血,分别在第1、2、4、6、8、10天用流式细胞仪检测。结果表明,肝素钠抗凝,第1、2、4天内检测差异不显著(P＞0.05),第6天检测CD4^＋、CD8^＋细胞显著减少(P＜0.05),第8、10天极显著减少(P＜0.01)。EDTA抗凝,第1、2、4天内检测差异不显著(P＞0.05),第6天检测CD3^＋细胞显著减少(P＜0.05),第8、10天极显著减少(P＜0.01);枸橼酸钠抗凝,第1、2、4、6、8天内检测差异不显著(P＞0.05),第10天检测CD3^＋、CD4^＋细胞极显著减少(P＜0.01),CD8^＋细胞显著减少(P＜0.05)。3种抗凝剂检测比较,处理后的第1、2、4、6天肝素钠抗凝管检测结果CD3^＋、CD4^＋、CD8^＋均高于EDTA和枸橼酸钠抗凝。本研究表明,流式细胞术检测猪外周血优选肝素钠抗凝,样本采集后4 d内检测完毕。

The Effects of T Lymphocyte Subset Detection for Flow Cytometry under Different Experimental Conditions

The experiments were designed to study the changed of T lymphocyte subset about CD3^＋,CD4^＋,CD8^＋ which detected by triple-color flow cytometry for swine,the blood sample were collected by three different anticoagulants and detection on different days. The results showed that CD3^＋,CD4^＋,CD8^＋ detected ratio on the 1st,2nd,4th day had no significant difference,but on the 6th CD4^＋,CD8^＋ had significant different,on the 8th day and 10th day had extremely significant different in the heparin anticoagulant samples group. In the EDTA anticoagulant samples group CD3^＋,CD4^＋,CD8^＋ detected ratio on the 1st,2nd and 4th day had no significant difference,on the 6th day CD3^＋ had significant difference,on the 8th and 10th day had extremely significant difference. In the sodium citrate anticoagulant samples group CD3^＋,CD4^＋,CD8^＋ detected ratio on the 1st,2nd,3rd,4th,6th day and the 8th day had no significant difference,on the 10th day CD3^＋,CD4^＋ had extremely significant difference,and CD8^＋ had significant difference.Compared three different anticoagulants,the heparin was better others,and it’s better to detect the samples within four days.