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A GFP-transfected HFWT cell line,GHINK-1, as a novel target for non-RI activated natural killer cytotoxicity assay
Authors:Hideki?Harada  Kaoru?Saijo  Isamu?Ishiwata  Email author" target="_blank">Tadao?OhnoEmail author
Institution:RIKEN Cell Bank, RIKEN (The Institute of Physical and Chemical Research), Koyadai 3-1-1, Tsukuba Science City, 305-0074, Japan.
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.
Keywords:Human activated NK cell  GFP  Cytotoxicity assay  HFWT cell line  non-RI
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