Clinical NK cell cryopreservation liquid and cryopreservation method

A technology of NK cells and cryopreservation method, which is applied in the field of clinical NK cell cryopreservation solution and cryopreservation, which can solve the problems of inability to meet the needs of mass production, limited sources of human autologous serum, and increased introduction of animal-derived components. Achieve the effect of maintaining the activity and killing ability of frozen cells, improving the activity and killing ability of frozen cells, and protecting mechanical damage

Pending Publication Date: 2021-02-26
COBAXER BIOTECH
View PDF7 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the dose of DMSO is large, it is harmful to the human body and cells. If you want to remove DMSO, you can only wash the frozen and recovered cells. Not only is the operation cumbersome, but also a large number of cells will be lost during the ope

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
  • Clinical NK cell cryopreservation liquid and cryopreservation method
  • Clinical NK cell cryopreservation liquid and cryopreservation method
  • Clinical NK cell cryopreservation liquid and cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046]Prepare NK cell cryopreservation solution for clinical use according to the following steps:

[0047]Step 1: Weigh the following components according to the volume ratio: glycerol 3%, dextran 15%, polyvinylpyrrolidone 0.1%, N-acetylcysteine ​​8%, glucose solution and half of water, and mix well to obtain A liquid;

[0048]Step 2: Mix DMSO and remaining water thoroughly to obtain liquid B.

[0049]Step 3: Store the solution A and solution B obtained in step 1 and step 2 at 2°C for 30 min in the dark.

[0050]Step 4: Mix the solution A and solution B obtained in step 3, add NK cells, and gently pipette until the density of the cell suspension is 8×107cells / ml; The process of NK cell processing is as follows: Centrifuge the NK cell suspension at 300g for 8 minutes, and discard the supernatant.

[0051]Step 5: Place the cell suspension in Step 4 in the dark at 4°C for 8 minutes, then cool it down, and transfer it to liquid nitrogen for storage.

[0052]Transfer to a program cooling box, place it at...

Embodiment 2

[0054]Prepare NK cell cryopreservation solution for clinical use according to the following steps:

[0055]Step 1: Weigh the following components according to the volume ratio: glycerol 3%, dextran 15%, polyvinylpyrrolidone 0.1%, N-acetylcysteine ​​8%, glucose solution and half of water, and mix well to obtain A liquid;

[0056]Step 2: Mix DMSO and remaining water thoroughly to obtain liquid B.

[0057]Step 3: Store the solution A and solution B obtained in step 1 and step 2 in the dark at 8°C for 30 minutes.

[0058]Step 4: Mix liquid A and B obtained in step 3, add NK cells, and gently pipette until the density of the cell suspension is 1×107cells / ml; The process of NK cell processing is as follows: Centrifuge the NK cell suspension at 300g for 8 minutes, and discard the supernatant.

[0059]Step 5: Place the cell suspension in Step 4 in the dark at 6°C for 8 minutes, then cool it down, and transfer it to liquid nitrogen for storage.

[0060]Transfer to a program cooling box, place it at -80°C, and...

Embodiment 3

[0062]Prepare NK cell cryopreservation solution for clinical use according to the following steps:

[0063]Step 1: Weigh the following components according to the volume ratio: glycerol 3%, dextran 15%, polyvinylpyrrolidone 0.1%, N-acetylcysteine ​​8%, glucose solution and half of water, and mix well to obtain A liquid;

[0064]Step 2: Mix DMSO and remaining water thoroughly to obtain liquid B.

[0065]Step 3: Store the solution A and solution B obtained in step 1 and step 2 at 6°C for 30 minutes in the dark.

[0066]Step 4: Mix the solution A and solution B obtained in step 3, add NK cells, and gently pipette until the density of the cell suspension is 8×107cells / ml; The process of NK cell processing is as follows: Centrifuge the NK cell suspension at 300g for 8 minutes, and discard the supernatant.

[0067]Step 5: Place the cell suspension in Step 4 in the dark at 4°C for 8 minutes, then cool it down, and transfer it to liquid nitrogen for storage.

[0068]Transfer to a program cooling box, place i...

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

PropertyMeasurementUnit
Cell densityaaaaaaaaaa
Login to view more

Abstract

The invention discloses a clinical NK cell cryopreservation solution and a cryopreservation method. The clinical NK cell cryopreservation solution comprises the following components in percentage by volume: 3%-10% of dimethyl sulfoxide, 2%-4% of glycerol, 13%-20% of dextran, 0.1%-0.2% of polyvinylpyrrolidone, 5%-10% of N-acetylcysteine and 60%-70% of a glucose solution. Wherein the glucose solution is formed by compounding medical-grade glucose and medical-grade pure water; the mass concentration of glucose in the glucose solution is 5wt%; the clinical NK cell cryopreservation solution can bedirectly used for intravenous injection, the DMSO content is low, the toxicity to human bodies or cryopreserved cells is greatly reduced, and the clinical NK cell cryopreservation solution is safer; the cell viability and killing capability can be better preserved, and the damage to the cells in the cryopreservation process is reduced.

Description

Technical field[0001]The present invention relates to the technical field of cell cryopreservation, in particular to a clinically used NK cell cryopreservation solution and a freezing storage method.Background technique[0002]Natural killer cell (Natural killer cell, NK) is a type of lymphocyte, derived from CD34+Hematopoietic progenitor cells achieve key immune regulation functions by regulating dendritic cells (DCs). As immune cells, NK cells have the function of resisting virus invasion and tumor killing. They are the body's first line of defense against tumors and have good application prospects in adoptive immunotherapy for malignant tumors. However, it occupies a very small comparison in peripheral blood, accounting for 5% to 15% of lymphocytes, and the function of NK cells in tumor patients is lacking in varying degrees, which makes it difficult to produce good anti-tumor effects. Therefore, NK cells have become one of the common types of immune cell therapy. At this stage, NK...

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
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0221A01N1/0226
Inventor 黄雨王晗程威谭莎李湘
Owner COBAXER BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap