Method of expanding nk cell and composition for culturing

Abstract

Provided are a method of ex-vivo culture of natural killer (NK) cells by treating the cells with a reactive oxygen species (ROS) inhibitor and/or a p53 protein inhibitor; and a composition comprising the cultured NK cells. By reducing the activity of ROS and p53 proteins during ex-vivo culture, NK cells may have achieved greater expansion efficiency without altering their anti-tumor cytotoxicity.

1 . A method of enhancing NK cell expansion, comprising:
 treating natural killer (NK) cells with one or more inhibitors selected from the group consisting of a reactive oxygen species (ROS) inhibitor and a p53 inhibitor.   
     
     
         2 . The method of  claim 1 , wherein NK cells are derived from human peripheral blood mononuclear cells (PBMCs).;I kn 
     
     
         3 . The method of  claim 1 , wherein the ROS inhibitor is one or more selected from the group consisting of 1-(4-hydroxy-3-methoxyphenyl)-ethanone (apocynin), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), pyrrolidine dithiocarbamate (PDTC), glutathione (GSH), catalase, manganese superoxide dismutase (MnSOD), vitamin E, and Quercetin. 
     
     
         4 . The method of  claim 1 , wherein the p53 inhibitor is one or more selected from the group consisting of 2-(2-imino-4,5,6,7-tetrahydrobenzothiazol-3-yl)-1-p-tolylethanone hydrobromide (pifithrin-α) and pifithrin-μ. 
     
     
         5 . The method of  claim 1 , wherein the inhibitors include trolox and pifithrin-α in a molar ratio of 50 to 150:1. 
     
     
         6 . The method of  claim 1 , wherein the treatment includes culturing NK cells expanded from PBMCs for 1 to 3 weeks in the presence of one or more inhibitor selected from the group consisting of the ROS inhibitor and the p53 inhibitor. 
     
     
         7 . The method of  claim 1 , wherein the method comprises:
 inducing and expending NK cells from PBMCs; and   culturing the expanded NK cells for 1 to 3 weeks in the presence of one or more inhibitor selected from the group consisting of the ROS inhibitor and the p53 inhibitor.   
     
     
         8 . A composition for culturing or storing natural killer (NK) cells, comprising one or more inhibitors selected from the group consisting of a reactive oxygen species (ROS) inhibitors and a p53 inhibitors as an active ingredient. 
     
     
         9 . The composition of  claim 8 , wherein the ROS inhibitor is one or more selected from the group consisting of 1-(4-hydroxy-3-methoxyphenyl)-ethanone (apocynin), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), pyrrolidine dithiocarbamate (PDTC), glutathione (GSH), catalase, manganese superoxide dismutase (MnSOD), vitamin E, and Quercetin. 
     
     
         10 . The composition of  claim 8 , wherein the p53 inhibitor is one or more selected from the group consisting of 2-(2-imino-4,5,6,7-tetrahydrobenzothiazol-3-yl)-1-p-tolylethanone hydrobromide (pifithrin-α) and pifithrin-μ. 
     
     
         11 . The composition of  claim 8 , wherein the composition includes trolox and pifithrin-α in a molar ratio of 50 to 150:1. 
     
     
         12 . A pharmaceutical composition in use for preventing or treating cancer, which comprises the natural killer (NK) cells obtained according to  claim 1 . 
     
     
         13 . A method of treating cancer, comprising:
 administering the natural killer (NK) cells obtained according to  claim 1  to a subject at an effective dose.