流式样本保存时间对肝癌患者外周血T淋巴细胞和NK细胞的影响

Effect of the storage time of samples for cytometry on peripheral blood T lymphocytes and NK cells of patients with hepatocellular carcinoma

目的观察样本保存时间对流式细胞仪检测肝癌患者外周血T淋巴细胞亚群的影响。方法随机选取肝癌患者16例,采血后6 h内分离外周血单核细胞,将细胞分成五组,包括立即上机组、4℃PBS保存1 d、2 d、3 d及5 d检测组,分别进行流式检测。结果样本保存3 d组淋巴细胞的物理参数发生变化;样本保存3 d组CD3+T淋巴细胞百分比显著降低(P <0. 05);样本保存3 d组NK细胞百分比显著降低(P <0. 05)。结论临床样本在收集后进行外周血单核细胞分离,将细胞用PBS于4℃条件下保存3 d内进行淋巴细胞和NK细胞的上机检测结果比较稳定。

Objective To study the effect of storage time of samples for cytometry on subsets of T lymphocytes of hepatocellular carcinoma patients. Methods Sixteen patients with hepatocellular carcinoma were collected randomly, and they were processed within 6 hours, then samples were tested by flow cytometry analysis within 1 hour, 1 day, 2 days, 3 days and 5 days, separately. The samples were stored in PBS at 4 ℃. Results At 3 days of samples treatment, the physical parameters of lymphocytes were changed;at 3 days of sample treatment, the percentage of surviving CD3 +T lymphocytes was decreased significantly ( P <0.05). At 3 days of sample treatment, the percentage of NK cells was decreased significantly ( P <0.05). Conclusion The results of T lymphocytes and NK cells of hepatocellular carcinoma patients analyzed by flow cytometry are stable within 3 days after the samples were processed stored in PBS at 4 ℃.