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流式细胞术中的自然杀伤细胞功能。I. 对K562细胞系的自然杀伤细胞裂解活性评估

Natural killer function in flow cytometry. I. Evaluation of NK lytic activity on K562 cell line.

作者信息

Papa S, Vitale M, Mariani A R, Roda P, Facchini A, Manzoli F A

机构信息

Istituto di Scienze Morfologiche, Urbino, Italy.

出版信息

J Immunol Methods. 1988 Feb 24;107(1):73-8. doi: 10.1016/0022-1759(88)90011-7.

Abstract

Flow cytometry has been employed to establish an NK assay using the K562 target cell line. These cells show a perpendicular light scatter (PLS) characteristically different from lymphocytes. Other physical parameters, such as forward light scatter (FLS), do not discriminate between the two populations, since dead K562 cells display similar FLS characteristics as effector cells. Killed cells are stained with propidium iodide and followed by flow analysis. It was advisable to add the dye in the medium so that, as long as the target cells are killed they will also be stained. Moreover the flow cytometric and trypan blue evaluation of target cell death rate showed a stronger correlation than did either test with the conventional 51Cr release assay, the first two methods both being based on the same biological mechanism.

摘要

流式细胞术已被用于建立一种使用K562靶细胞系的自然杀伤细胞(NK)检测方法。这些细胞表现出与淋巴细胞特征性不同的垂直光散射(PLS)。其他物理参数,如前向光散射(FLS),无法区分这两种细胞群体,因为死亡的K562细胞与效应细胞表现出相似的FLS特征。死亡细胞用碘化丙啶染色,然后进行流式分析。建议在培养基中加入染料,这样只要靶细胞被杀死,它们也会被染色。此外,流式细胞术和台盼蓝对靶细胞死亡率的评估显示出比传统的51Cr释放试验更强的相关性,前两种方法都基于相同的生物学机制。

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