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Immune-cell in-vitro-induction-amplification method

A technology of immune cells and nuclear cells, which is applied in the direction of blood/immune system cells, animal cells, cell culture medium, etc. It can solve the problems of natural killer cell culture methods that need to be improved, poor amplification multiples and tumor killing effects, and complicated culture systems And other issues

Active Publication Date: 2017-05-31
ACADEMY OF MILITARY MEDICAL SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, NK cell therapy products currently on the market have problems such as complicated culture systems, poor amplification multiples, and poor tumor killing effects, and some culture systems have safety risks.
[0004] Therefore, the culture method of natural killer cells needs to be improved

Method used

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Embodiment 1

[0052] Using the immune cell culture medium system of the embodiment of the present invention, the isolated mononuclear cells are induced and expanded into immune cells, and the activity of the immune cells is detected.

[0053] 1. Experimental method

[0054] 1. Prepare anti-human CD16 coated T75 vials

[0055] 1.1 Add 5 mL of 2.5 μg / mL anti-human CD16 monoclonal antibody dissolved in medical normal saline into a sterile culture bottle, shake the culture bottle gently to make the antibody cover the culture surface, and overnight at 4°C in the dark.

[0056] 1.2 Recover the antibody coating solution before use, wash the culture bottle once with 5 mL of normal saline, and then use 5 mL of T cell expansion medium (OpTmizer TM CTS TM T-cell expansion SFM) and washed once.

[0057] 2. Collect peripheral blood, separate peripheral blood plasma and mononuclear cells

[0058] 2.1 Collect about 100ml of human peripheral blood with a sterile blood collection bag added with antico...

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Abstract

The invention discloses an immune-cell in-vitro-induction-amplification method. The immune-cell in-vitro-induction-amplification method includes the steps that a culturing container is coated with a CD16 antibody to obtain the coated culturing container; media are activated through immune cells, and mononuclear cells are put into the coated culturing container for activating the media through the immune cells to obtain activated immune cells; the media are amplified through the immune cells, the immune cells subjected to primary induction amplification are subjected to secondary induction-amplification culturing to obtain amplified immune cells; the media are amplified in a large-scale mode through the immune cells, differentiated immune cells are subjected to tertiary induction-amplification culturing to obtain large-scale-amplified functional activated immune cells. The mononuclear cells are subjected to induction-amplification culturing with the immune-cell in-vitro-induction-amplification method to obtain the large-scale immune cells, the immune-cell in-vitro-induction-amplification method has the advantages of being high in induction efficiency, amplification speed and safety, low in cost and the like, and therefore the requirements of a large number of the immune cells in clinic treatment are met.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing and expanding immune cells in vitro. Background technique [0002] In recent years, biological therapy has become the fifth largest treatment mode after surgery, radiotherapy and chemotherapy, and endocrine therapy, and has gradually received attention. Adoptive cellular immunotherapy (ACI) is one of the methods of cell biological therapy. It refers to the infusion of immune cells with anti-tumor activity to tumor patients, directly killing tumor cells or stimulating the body's immune response to kill tumor cells, so as to achieve the goal of treating tumors. Purpose. Currently, adoptive immune cells in clinical use include DC-CIK cells, TIL cells, LAK cells and NK cells, among which CIK cells, LAK cells and A-NK cells are all anti-cancer systems with natural killer cells as the main body. [0003] Natural killer cells (natural killer cells) are also called NK...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0037C12N5/0646C12N2500/90C12N2501/06C12N2501/2302C12N2501/599
Inventor 裴雪涛习佳飞岳文陈琳姚海雷南雪张亚
Owner ACADEMY OF MILITARY MEDICAL SCI
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