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NK cell in-vitro amplification composition and NK cell amplification method

A NK cell, in vitro expansion technology, applied in the fields of botanical equipment and methods, receptors/cell surface antigens/cell surface determinants, biochemical equipment and methods, etc. The problems of long cycle and low purity can shorten the screening cycle, reduce production costs and avoid pollution.

Pending Publication Date: 2016-10-05
天晴干细胞股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to solve the problems of low amplification factor, low purity, long amplification period and high cost of NK cells at present, and provides a NK cell in vitro amplification composition and an NK cell expansion method

Method used

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  • NK cell in-vitro amplification composition and NK cell amplification method
  • NK cell in-vitro amplification composition and NK cell amplification method
  • NK cell in-vitro amplification composition and NK cell amplification method

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specific Embodiment approach 1

[0041] Specific embodiment 1: The NK cell expansion composition in this embodiment includes NK expansion trophoblast cells, IL-2, inactivated autologous plasma and GT-T551 H3 medium, wherein each milliliter of GT-T551 H3 medium is added 0.65×10 6 ~1×10 6 NK expansion trophoblasts, add 200-500IU of IL-2 to every milliliter of GT-T551 H3 medium, the volume of inactivated autologous plasma is 5% of the volume of GT-T551 H3 medium, the NK expansion trophoblasts The cells are K562-4-1BBL-IL-15-IL-21 trophoblasts.

[0042] The in vitro expansion method of NK cells in the present invention is mainly to inlay factors that promote the activation and proliferation of NK cells on the surface of K562 cells that have an inductive effect on NK cells, so that NK cells can be better activated and expanded, so that they have Better tumor cell killing activity; this method takes less time and has high safety, and can be applied to clinical cell biological therapy, including renal cancer, mali...

specific Embodiment approach 2

[0043] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that 0.7×10 6 ~0.9×10 6 NK expansion trophoblasts. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0044] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: adding 0.8×10 6 NK expansion trophoblasts. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses an NK cell in-vitro amplification composition and an NK cell amplification method and relates to an NK cell amplification composition and an amplification method. The invention aims at solving the problems of low multiple, low purity, long amplification period and high cost of current NK cell amplification. The composition comprises NK amplification trophocyte, IL-2, inactivated autologous plasma and GT-T551 H3 culture medium. The amplification method comprises the following steps: re-suspending PBMCs with the GT-T551 H3 culture medium; transferring into a culture bottle; adding the IL-2, NK amplification trophocyte and autologous plasma; and performing in-vitro co-culture for 15 days, wherein the NK amplification trophocyte is supplemented in the 7th day of the culture, the culture medium is supplemented in the culture process, and the cells are collected for detection in the 13th and 15th days of the culture. The composition can remarkably promote mass amplification of NK cells to meet the needs of clinical application; and the obtained NK cells have relatively high purity, the cost is saved, and the killing activity is improved. The NK cell in-vitro amplification composition and NK cell amplification method are applied to the amplification of NK cells.

Description

technical field [0001] The invention relates to an NK cell expansion composition and an expansion method. Background technique [0002] The latest edition of the "World Cancer Report" comprehensively describes and analyzes the overall situation and prevalence trends of 28 cancers in more than 180 countries around the world. The report predicts that global cancer cases will show a rapid growth trend. 14 million cancer cases and 8.2 million deaths. Among them, China added 3.07 million cancer patients and caused about 2.2 million deaths, accounting for 21.9% and 26.8% of the global total respectively. Among them, lung cancer, gastric cancer, and liver cancer are the cancers with the highest incidence rates, while breast cancer, colorectal cancer, and cervical cancer threaten women's health. [0003] Tumor immunotherapy is recognized as the fourth major tumor treatment method by domestic and foreign medical circles, among which the autologous immune cell (T cell, NK cell) trea...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12N5/10C12N15/867C12N7/01
CPCC12N5/0646C07K14/54C07K14/5443C07K14/70578C07K2319/02C07K2319/03C12N7/00C12N15/86C12N2501/2302C12N2502/30C12N2510/00C12N2740/15021C12N2740/15043
Inventor 张怡刘春香芦慧颖李琳王浩宇车彦川刘艳青
Owner 天晴干细胞股份有限公司
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