Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

CAR-NK cell, and preparation method and application thereof

A kind of NK cell and cell technology, applied in the direction of biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problem that the anti-tumor function cannot be fully exerted, and achieve the effect of a healthy fetus with a smooth labor process and no history of infectious diseases

Active Publication Date: 2019-03-29
WUHAN BIO RAID BIOTECH CO LTD
View PDF3 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although tumor cells have lost classic HLA-class I molecules, the expression of non-classic HLA-class I molecules HLA-G, HLA-E, etc. can also inhibit the activation of NK cells
At the same time, due to the decline in the number and quality of NK cells in tumor patients and the existence of tumor escape mechanisms, their anti-tumor functions in vivo have not been fully exerted.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • CAR-NK cell, and preparation method and application thereof
  • CAR-NK cell, and preparation method and application thereof
  • CAR-NK cell, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Cord Blood Mononuclear Cell Extraction

[0046] (1) Take 50 mL of cord blood, divide it into two 50 mL centrifuge tubes, centrifuge at 650 g for 15 min at room temperature, take the upper layer of light yellow plasma into a new 50 mL centrifuge tube (the lower layer of red liquid is used to extract mononuclear cells), The plasma was inactivated in a water bath at 56°C for 30min, then centrifuged at 900g for 10min, the supernatant was taken, and placed in an environment of -20°C for 15min; centrifuged again at 900g, 10min, the supernatant was taken and stored at 4°C until use. (The centrifuge adjusts the speed up by 1 and the speed down by 1).

[0047] (2) Take the lower red liquid obtained in the previous step of plasma extraction and dilute it with an equal volume of normal saline to a total volume of about 20 mL, mix it upside down, and set aside. Take another 2 new 50mL centrifuge tubes, and add 20mL of lymphocyte separation solution to each tube. Add 20m...

Embodiment 2

[0049] Example 2 Induction and activation of umbilical cord blood NK cells

[0050] The mononuclear cells obtained in Example 1 were divided into 2 × 10 6 Individuals / mL inoculated with 1-10% cord blood autologous plasma, 200-1500 IU / mL IL-2, 10-50 ng / mL IL-15, 10-30 ng / mL IL-7 and 50-100 ng / mL IL-21 lymphocyte culture medium, placed at 37 °C, 5% CO 2 Cultured in an incubator, and on the 3rd day according to the cell density of 0.8~1.0×10 6 pc / mL rehydration solution.

Embodiment 3

[0051] Example 3 Detection of NK cell surface markers derived from peripheral blood and umbilical cord blood

[0052] Take 50 mL of peripheral blood, extract mononuclear cells according to the method in Example 1, and carry out activation culture according to the method in Example 2.

[0053]Take the cells in Example 2 and the activated cultured NK cells derived from peripheral blood, wash them twice with PBS, then add mouse IgG, and keep away from light for 30 minutes at 4°C; add specific antibodies CD16, CD161, NKG2A, NKG2D, NKp46, respectively. Incubate at ℃ for 30 min in the dark. The stained cells were washed twice, and the above markers were detected by flow cytometry. Statistical methods using SPSS10 statistical software. All data are expressed as x±s, and t test and Mann-Whitney test were used. The results are shown in Table 1. There was no significant difference in phenotype between the two sources of NK cells.

[0054] Table 1 Expression of NK cell surface marker...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a CAR-NK cell, and a preparation method and application thereof, and belongs to the field of cell therapy and biopharmaceutics. The method uses umbilical cord blood as a sourceof NK cells, and solves the problems of CAR-NK cells from a NK cell line, peripheral blood and autologous blood, such as complicated operation, risk of tumor formation, insufficient cell activity, asmall number of cells and GVHD caused by partial allogeneic origin cells.

Description

technical field [0001] The invention belongs to the field of cell therapy and the field of biopharmaceuticals, and specifically relates to CAR-NK cells using umbilical cord blood as a source of allogeneic cells, and using human chimeric antigen receptor modification to treat tumors and infectious diseases. Background technique [0002] Chimeric antigen receptor (CAR)-modified T cells (CAR-T) have made a major breakthrough in leukemia research, bringing new hope to patients with blood cancers. However, CAR-T therapy also has some problems, such as off-target effects, cytokine storms, insertion mutations, etc., and has not yet achieved significant curative effect on solid tumors. The development of new effector cells with strong anti-tumor effect has important theoretical significance and clinical application value. NK cells (natural killer cells, natural killer cells) are considered to have the potential to enhance their anti-tumor ability through CAR modification because of...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10A61K35/17A61P35/00A61P31/00
CPCA61K35/17A61P31/00A61P35/00C07K14/7051C12N5/0646C12N2510/00
Inventor 张同存顾潮江周勇
Owner WUHAN BIO RAID BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com