小鼠脾脏淋巴细胞体外培养前后细胞表面标记物CD3、CD4、CD44、CD62L 的变化

目的:探讨体外培养小鼠脾脏淋巴细胞后,细胞表面标记物CD3、CD4、CD44、CD62L 的变化。方法:用淋巴细胞分离液分离小鼠脾脏淋巴细胞,37益细胞孵箱中体外培养3 d 后,向细胞中加入流式抗体Mouse CD3e PE-CY7、Mouse CD4 FITC、Mouse CD44 PE、Mouse CD62L APC,同时设阴性对照,应用流式细胞仪检测细胞表面标记物CD3、CD4、CD44、CD62L 的水平。结果:小鼠脾脏淋巴细胞分离后直接做流式检测,CD3+ CD4+细胞占19.09%,其中98.61% 的细胞为CD44+ ,68.71% 为CD62L+ 。体外培养后CD3+ CD4+细胞占8.96%,其中71.82%为CD44+ ,11.27%为CD62L+ 。哮喘小鼠脾脏淋巴细胞分离后直接做流式检测,CD3+ CD4+ 细胞占20.33%,其中97.72% 的细胞为CD44+ ,75.74% 为CD62L+ 。体外培养后CD3+ CD4+ 细胞占7.2%,CD44 阳性细胞占58.21%,CD62L 阳性细胞仅占2.77%。结论:小鼠脾脏淋巴细胞体外培养后,细胞表面标记物CD44和CD62L 发生巨大变化,CD62L 呈现明显下调趋势,接近消失,CD44 也有下调。

Changes of cell surface markers CD3,CD4,CD44,CD62L on mouse spleen lym-phocytes after culture in vitro

Objective:To study the changes of cell surface markers CD3,CD4,CD44 and CD62L on mouse spleen lymphocytes after culture in vitro.Methods: Spleen cells were separated from BALB/ c mice and asthmatic mice and lymphocytes were isolated by using lymphocyte separation liquid.We cultured lymphocytes with RPMI1640 medium(10% FBS) in cell incubator for 3 days.Then the four cell surface markers were detected by using flow cytometry.Results: In normal mice,19.09% of lymphocytes stained positively for CD3 and CD4,among these cells 98.61% was CD44+ T cell and 68.71% was CD62L+ T cell before the culture.After culture with ConA for 3 days,CD3+ CD4+ and CD3+ CD4+ CD44+ T cell population was down regulated to 8.96% and 71.82%,respectively.While CD62L almost disappeared, accounting for 11.27% only.For asthmatic mice, CD3+ CD4+ T cell accounted for 20.33% in total lymphocytes,among which 97.72% was CD44+ and 75.74% was CD62L+ before the culture.After culture,CD3+ CD4+ T cell accounted for 7.2%,among which CD44+ only 58.21% and CD62L+ only 2.77%.Conclusion: In both normal and asthmatic mice,CD3,CD4,CD44 and CD62L on spleen lymphocytes were down regulated after culture in vitro regardless with or without stimulating factors,especially CD62L,almost disappeared.This should be given full consideration when these markers are detected after cell culture.