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
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摘要
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 ℃.
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 ℃.
引文
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[8]方芳,陈柯霖,刘淑静,等.标本保存时间及温度对流式细胞仪检测非健康人外周血淋巴细胞亚群影响研究[J].检验医学与临床,2016,13(9):1164-1166. DOI:10. 3969/j. issn. 1672-9455. 2016.09. 005.FANG F,CHEN K L,LIU S J,et al. The influence of storage timeand temperature on flow cytometry analysis of patients'peripheral bloodlymphocyte subsets[J]. Lab Med Clin,2016,13(9):1164-1166.DOI:10. 3969/j. issn. 1672-9455. 2016. 09. 005.
[2]王建中,王淑娟.当前临床流式细胞分析的发展趋势[J].中华检验医学杂志,2002,25(1):5-7. DOI:10. 3760/j:issn:1009-9158.2002. 01. 001.
[3] OKADA Y. Flow cytometry--the basis of cell surface analysis and clini-cal application[J]. Rinsho Byori,2010,58(11):1121-1130.
[4] AVLASEVICH S,BRYCE S,DE BOECK M,et al. Flow cytometricanalysis of micronuclei in mammalian cell cultures:past,present andfuture[J]. Mutagenesis,2011,26(1):147-152. DOI:10. 1093/mutage/geq058.
[5] SIEBERT J C,WALKER E B. Monitoring cytokine profiles during im-munotherapy[J]. Immunotherapy,2010,2(6):799-816. DOI:10.2217/imt. 10. 76.
[6]陈潇,吴丽娟.抗凝剂、标本保存温度和保存时间对流式T淋巴细胞亚群检测的影响[J].国际检验医学杂志,2012,33(2):146-147,150. DOI:10. 3969/j. issn. 1673-4130. 2012. 02. 008.CHEN X,WU L J. Empirical study of the influence of decoagulant,storage temperature and time on the detection of peripheral bloodT-lymphocyte subgroups by flow cytometry[J]. Int J Lab Med,2012,33(2):146-147,150. DOI:10. 3969/j. issn. 1673-4130. 2012.02. 008.
[7] EKONG T,KUPEK E,HILL A,et al. Technical influences on immu-nophenotyping by flow cytometry. The effect of time and temperature ofstorage on the viability of lymphocyte subsets[J]. J ImmunolMethods,1993,164(2):263-273.
[8]方芳,陈柯霖,刘淑静,等.标本保存时间及温度对流式细胞仪检测非健康人外周血淋巴细胞亚群影响研究[J].检验医学与临床,2016,13(9):1164-1166. DOI:10. 3969/j. issn. 1672-9455. 2016.09. 005.FANG F,CHEN K L,LIU S J,et al. The influence of storage timeand temperature on flow cytometry analysis of patients'peripheral bloodlymphocyte subsets[J]. Lab Med Clin,2016,13(9):1164-1166.DOI:10. 3969/j. issn. 1672-9455. 2016. 09. 005.