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一种绿色荧光蛋白(GFP)转染的HFWT细胞系GHINK-1,作为非RI激活的自然杀伤细胞细胞毒性测定的新靶点。

A GFP-transfected HFWT cell line, GHINK-1, as a novel target for non-RI activated natural killer cytotoxicity assay.

作者信息

Harada Hideki, Saijo Kaoru, Ishiwata Isamu, Ohno Tadao

机构信息

RIKEN Cell Bank, RIKEN (The Institute of Physical and Chemical Research), Koyadai 3-1-1, Tsukuba Science City, 305-0074, Japan.

出版信息

Hum Cell. 2004 Mar;17(1):43-8. doi: 10.1111/j.1749-0774.2004.tb00019.x.

Abstract

An anchorage-dependent Wilms' tumor cell line, HFWT, has been found to be extremely susceptible to human natural killer (NK) cells. Here we established a transfectant of HFWT with the green fluorescence protein (GFP) gene, designated GHINK-1 cells, to apply to the activated NK cytotoxicity assay without radioisotope labeling. After being co-cultured with CD3 CD56+ NK cells, GHINK-1 cells released GFP into the medium. The intensity of the fluorescence from the released GFP correlated almost exactly with the radioactivity of a standard 51Cr-release assay performed with suspension-cultured K562 cells. Because it does not require separation of the remaining live target cells by centrifugation, the non-radioisotopic GFP release assay with GHINK-1 cells is a convenient alternative for monitoring human activated NK killing activity.

摘要

一种锚定依赖型肾母细胞瘤细胞系HFWT,已被发现对人类自然杀伤(NK)细胞极为敏感。在此,我们构建了一个转染了绿色荧光蛋白(GFP)基因的HFWT转染细胞系,命名为GHINK-1细胞,用于无放射性同位素标记的活化NK细胞毒性检测。与CD3 CD56+ NK细胞共培养后,GHINK-1细胞将GFP释放到培养基中。释放出的GFP荧光强度与用悬浮培养的K562细胞进行的标准51Cr释放检测的放射性几乎完全相关。由于该检测不需要通过离心分离剩余的活靶细胞,因此使用GHINK-1细胞进行的非放射性同位素GFP释放检测是监测人类活化NK细胞杀伤活性的一种便捷替代方法。

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