US 11,841,369 B2
Method for improving quality of therapeutic cell through real-time glutathione measurement
Heun Soo Kang, Seoul (KR); Hye Mi Kim, Seoul (KR); Ji Eun Song, Seoul (KR); Gwang Mo Yang, Seoul (KR); Ji Woong Shin, Seoul (KR); Hye Won Kang, Seoul (KR); Yong Hwan Kim, Gyeonggi-do (KR); and Myung Jin Kim, Seoul (KR)
Assigned to Cell2In, Inc., Seoul (KR)
Appl. No. 16/767,985
Filed by CELL2IN, INC., Seoul (KR)
PCT Filed Nov. 28, 2018, PCT No. PCT/KR2018/014815
§ 371(c)(1), (2) Date May 28, 2020,
PCT Pub. No. WO2019/107913, PCT Pub. Date Jun. 6, 2019.
Claims priority of application No. 10-2017-0160563 (KR), filed on Nov. 28, 2017; and application No. 10-2018-0094878 (KR), filed on Aug. 14, 2018.
Prior Publication US 2021/0003582 A1, Jan. 7, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/68 (2006.01); G01N 21/64 (2006.01); G01N 33/50 (2006.01); A61K 38/06 (2006.01)
CPC G01N 33/68 (2013.01) [A61K 38/063 (2013.01); G01N 21/6428 (2013.01); G01N 33/5005 (2013.01); G01N 33/5044 (2013.01); G01N 33/5076 (2013.01); G01N 33/6815 (2013.01); G01N 33/6893 (2013.01); G01N 2021/6439 (2013.01); G01N 2800/7009 (2013.01)] 9 Claims
 
1. A method of improving the quality of cells, comprising:
isolating desired cells;
measuring a glutathione (GSH) level in the isolated cells;
determining cell quality according to the GSH level,
wherein the determination of cell quality according to the GSH level is performed based on one or more GSH evaluation parameters selected from the group consisting of:
i) glutathione regeneration capacity (GRC) of the isolated cells, wherein GRC is a value obtained by real-time monitoring of FR or F510 after the isolated cells are treated with an oxidizing agent, and is calculated by dividing a value obtained by subtracting a second area under the curve (AUC) of a group of the isolated cells treated with a second oxidizing agent from a first AUC of a group of the isolated cells treated with a first oxidizing agent by a value obtained by subtracting the second AUC of the group of the isolated cells treated with the second oxidizing agent from a third AUC of a naive control and multiplying the resulting value by 100, wherein FR is an fluorescence emission ratio of a fluorescence emission value at 510 nm (F510) to a fluorescence emission value at 580 nm (F580); and
ii) oxidative stress resistance capacity (ORC) of the isolated cells, wherein ORC is a value of cell counts with a variation in GSH expression, obtained by comparing a GSH level quantified after the isolated cells are treated with a first oxidizing agent with a GSH level quantified in control cells which are not treated with an oxidizing agent or in control cells which have not yet been treated with an oxidizing agent; and
adding a material capable of improving the one or more GSH evaluation parameters into the isolated cells, wherein the material is any one or more selected from the group consisting of glutathione ethyl ester, ascorbic acid 2-glucoside, glutathione, N-acetylcysteine, 2-mercaptoethanol, dithiothreitol (DTT), cysteine, γ-glutamyl cysteine (GGC), GGC esters, oxo-4-thiazolidinecarboxylic acid (OTC), L-2-oxo-4-thiazolidinecarboxylic acid, lipoic acid, Ferrostatin-1, Liproxstatin-1, vitamin D3, 1-alpha, 25-dihydroxy VitD3, vitamin E, coenzyme Q10, and an iron or copper ion chelator selected from the group consisting of deferoxamine, deferiprone and deferasirox, baicalin, baicalein, luteolin, quercetin, butein, flower extracts of Chrysanthemum morifolium Ramat, leaf extracts of Cedela sinensis A. Juss, extracts of Oenothera stricta Ledeb., extracts of Equisetum arvense L., leaf extracts of Ipomoea batatas, tomato extracts, and homocysteine.