NK cell quantity amplification method based on expression of surface-activated receptors and inhibition receptors

A technology of NK cells and surface activation, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of small number of replacements, inability to convert, inconvenient cell observation, etc., to increase the quality of cell culture, improve The effect of convenience

Pending Publication Date: 2021-10-22
AFFILIATED CANCER HOSPITAL OF SHANDONG FIRST MEDICAL UNIV SHANDONG CANCER INST (SHANDONG CANCER HOSPITAL)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 1. In the existing NK cell number expansion research, in most research experiments, the vessels used are mostly fixed, and the number of replacements is small, which cannot be converted according to the changes, and it is inconvenient to observe the cells;
[0004] 2. After the research, it is impossible to conduct research based on the addition of various inducing factors, and it is impossible to determine the specific effects of NK cells caused by different factors

Method used

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  • NK cell quantity amplification method based on expression of surface-activated receptors and inhibition receptors
  • NK cell quantity amplification method based on expression of surface-activated receptors and inhibition receptors
  • NK cell quantity amplification method based on expression of surface-activated receptors and inhibition receptors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] see Figure 1-Figure 2 A method for expanding the number of NK cells based on the expression of surface-activated receptors and inhibitory receptors, comprising the steps of:

[0042] S1: Preparation of raw materials: autologous or allogeneic NK cells are used for transplantation, and cells can be obtained from different sources, including peripheral blood mononuclear cells, umbilical cord blood, bone marrow, induced pluripotent stem cells and cell lines, and multiple groups are used for culturing experiments appliance;

[0043] S2: Experiment: expand NK cells in vitro, use cytokines to expand NK cells and increase their activity, cytokines mediate the proliferation and activation of NK cells, and the cytokines include IL-2, IL-15, IL-12 , IL-18, IL-21 and type I interferon;

[0044] S3: Expansion observation: place the taken out raw materials in the culture vessel and add inducing amplification factors for mixing, and conduct continuous observation of the cells in th...

Embodiment 2

[0063] The experiment for S2 includes the following steps:

[0064] S201: Take one of the peripheral blood mononuclear cells, umbilical cord blood, bone marrow, induced pluripotent stem cells and cell lines, after passing the virus test, send it to the laboratory, take 70ml of blood cells, and separate them according to the conventional plasma and PBMC separation methods Obtain plasma and buffy coat;

[0065] S202: After washing the white blood cell layer, the count is 7.4*107, inoculated in the culture vessel at a density of 1.5*106 / ml, take multiple groups of cell bodies for detection, and place them in different culture vessels for amplification experiments ;

[0066] S203: Add induction medium, IL-2 and IL-15 co-factors and inactivated plasma, and culture in a carbon dioxide incubator at 37°C and 5% CO2.

[0067]The blood cells are collected as fresh blood, and cord blood is collected as fresh blood as possible. The specific gravity of the layered liquid for separating h...

Embodiment 3

[0069] For the experiment in S2, the following steps are included:

[0070] S201: Take one of the peripheral blood mononuclear cells, umbilical cord blood, bone marrow, induced pluripotent stem cells and cell lines, after passing the virus test, send it to the laboratory, take 70ml of blood cells, and separate them according to the conventional plasma and PBMC separation methods Obtain plasma and buffy coat;

[0071] S202: After washing the white blood cell layer, the count is 7.4*107, inoculated in the culture vessel at a density of 1.5*106 / ml, take multiple groups of cell bodies for detection, and place them in different culture vessels for amplification experiments ;

[0072] S203: Add induction medium, IL1-12, IL-15 and IL-18 combined factors and inactivated plasma, and place in a carbon dioxide incubator at 37°C and 5% CO2 for cultivation.

[0073] The blood cells are collected as fresh blood, and cord blood is collected as fresh blood as possible. The specific gravity ...

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Abstract

The invention discloses an NK (Natural Killer) cell quantity amplification method based on expression of surface-activated receptors and inhibition receptors, and belongs to the technical field of NK cell quantity amplification. The NK cell quantity amplification method comprises the steps five steps of raw material preparation, experiment performing, amplification observation, cell detection and conclusion comparison. According to the NK cell quantity amplification method based on expression of the surface-activated receptors and the inhibition receptors, more sufficient oxygen exchange is provided for cells, nutrients with larger sources can be provided for the cells, a good convection environment is provided for the cells, solute in the culture medium can freely move to the cells through natural convection, and the culture convenience is improved. And the cell culture quality is improved, and an amplification scheme can be further discussed and optimized, so that the efficient and high-purity NK cells are obtained, later browsing and research on experimental data are facilitated, foundation is prepared for next research, and the later research quality is improved.

Description

technical field [0001] The invention relates to the technical field of NK cell number expansion, in particular to a NK cell number expansion method based on the expression of surface activated receptors and inhibitory receptors. Background technique [0002] Natural killer cells (NK cells) play an important role in the body's innate immune system. Researchers described them as early as the 1970s, but it was not until the last 15 years that people gradually understood NK cells. The complexity and therapeutic potential of cells to help fight cancer and other diseases. [0003] 1. In the existing NK cell number expansion research, in most research experiments, the vessels used are mostly fixed, and the number of replacements is small, which cannot be converted according to the changes, and it is inconvenient to observe the cells; [0004] 2. After the research, it is impossible to conduct research based on the addition of various inducing factors, and it is impossible to deter...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2302C12N2501/2315C12N2501/2312C12N2501/2318C12N2501/2321C12N2501/24
Inventor 孟祥姣黄召勤李慧娟彭建峰
Owner AFFILIATED CANCER HOSPITAL OF SHANDONG FIRST MEDICAL UNIV SHANDONG CANCER INST (SHANDONG CANCER HOSPITAL)
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