外周血T淋巴细胞的培养、纯化及鉴定

Cultivation, purification and identification of T lymphocytes from peripheral blood

目的:建立外周血T淋巴细胞初步纯化方法。方法:取健康人外周血单个核细胞(PBMCs),在低剂量IL2维持下,利用细胞培养淘汰法筛选纯化T淋巴细胞,用流式细胞术检测淋巴细胞培养过程中细胞表面标志的改变。结果:用流式细胞术检测发现:B淋巴细胞于第1周时基本死亡,第3周时完全消失;NK细胞于第1周时明显增加,第4周时大部分死亡;第4周时,所培养的细胞绝大多数为T淋巴细胞。T淋巴细胞亚群的整体变化趋势为:随着培养时间的延长,CD4+百分数稍增加,CD8+百分数明显增加,CD4+/CD8+比值明显降低,CD3+CD16+CD56+百分数明显增加。结论:在IL2维持下,人PBMCs培养4周时,收获培养细胞为初步纯化的T淋巴细胞。

Aim： To establish the methods of purifying T lymphocytes from peripheral blood. Methods： With flow cytometry （FCM） , the volunteer＇s peripheral blood mononuclear cells （PBMCs） co-cultivated with IL-2 were observed to get their differentiation information, Results： Most of B lymphocytes died in the first week of culture and none of them were detected in the third week. The natural killer cells （NK） increased in the first week and most of them died in the fourth week. The CD4^＋/CD8^＋ ratio decreased obviously, and the percent of CD3^＋ CD16 ^＋ CD56 ^＋ increased obviously in the meantine. Conclusion ： Most of the PBMCs cultivated with IL-2 in vitro are initial purified T lymphocytes in the fourth week.