Unlock instant, AI-driven research and patent intelligence for your innovation.

A recovery method for car-t cells

A cell, 5% CO2 technology, applied in the field of CAR-T cell recovery, can solve the problems of cryopreservation tube explosion, long recovery time of viability, low recovery of CAR-T cells, etc. The effect of reducing the risk of explosion

Active Publication Date: 2020-04-03
SICHUAN HUIYU PHARMA
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, at present, the recovery rate of CAR-T cells is low, the recovery time of the recovery rate is long, and there is a risk of explosion of cryopreserved tubes when recovering CAR-T cells

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
  • A recovery method for car-t cells
  • A recovery method for car-t cells
  • A recovery method for car-t cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] After taking out a CAR-T cell cryopreservation tube from the liquid nitrogen tank, place it at room temperature at 20°C for about 30 seconds; place the cryopreservation tube in a 40°C water bath to thaw for 120 seconds, and ensure that the mouth of the cryopreservation tube is above the liquid surface. Gently shake the cryopreservation tube to thaw it quickly; after thawing, transfer the cell solution in the cryopreservation tube to a centrifuge tube containing 9ml CAR-T cell growth medium in a biological safety cabinet, 800rpm, Centrifuge for 5 minutes, discard the supernatant, add growth medium to gently resuspend the cells, transfer to a square bottle, leave 0.5ml of cell fluid for trypan blue staining, and count by hemocytometer method, the counting result is 4.3×10 5 cells / ml, cell viability 88.7%; place the square bottle on a plane cell shaker placed in a carbon dioxide incubator, 37°C, 5% CO 2 , saturated humidity, 50mm amplitude, 15rpm, micro-dynamic culture, cu...

Embodiment 2

[0034]After taking out a CAR-T cell cryopreservation tube from the liquid nitrogen tank, place it at room temperature at 25°C for about 20 seconds; place the cryopreservation tube in a 45°C water bath to thaw for 60 seconds, and ensure that the mouth of the cryopreservation tube is above the liquid surface. Gently shake the cryopreservation tube to thaw it quickly; after thawing, transfer the cell solution in the cryopreservation tube to a centrifuge tube containing 9ml CAR-T cell growth medium in a biological safety cabinet, 900rpm, Centrifuge for 4 minutes, discard the supernatant, add growth medium to gently resuspend the cells, transfer to a square bottle, leave 0.5ml of cell fluid for trypan blue staining and counting, and the counting result is 4.3×10 5 cells / ml, cell viability 87.6%; place the square bottle on a plane cell shaker placed in a carbon dioxide incubator, 37°C, 5% CO 2 , saturated humidity, 25mm amplitude, 25rpm, micro dynamic culture, cultured for 76h, afte...

Embodiment 3

[0039] After taking out a CAR-T cell cryopreservation tube from the liquid nitrogen tank, place it at room temperature at 15°C for about 60 seconds; place the cryopreservation tube in a water bath at 39°C for 180 seconds, and ensure that the mouth of the cryopreservation tube is above the liquid surface. Gently shake the cryopreservation tube to thaw it quickly; after thawing, transfer the cell solution in the cryopreservation tube to a centrifuge tube containing 9ml CAR-T cell growth medium in a biological safety cabinet, 1000rpm, Centrifuge for 3 minutes, discard the supernatant, add growth medium to gently resuspend the cells, transfer to a square bottle, leave 0.5ml of cell fluid for trypan blue staining and counting, and the counting result is 4.4×10 5 cells / ml, cell viability 87.6%; place the square bottle on a plane cell shaker placed in a carbon dioxide incubator, 37°C, 5% CO 2 , saturated humidity, 75mm amplitude, 5rpm, micro-dynamic culture, cultured for 72h, after t...

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 relates to the field of cell culture, in particular to a CAR-T cell resuscitation method. According to the resuscitation method, the average resuscitation rate of CAR-T cells is increased to 85% or above; the exploding risk of cryogenic vials during CAR-T cell resuscitation is greatly lowered; the CAR-T cell motility rate recovery is accelerated; micro-dynamic culture is performed at the amplitude being 25-75 mm, the rotating speed being 5-25 rpm, the temperature being 37 DEG C, the CO2 concentration being 5% and the saturation humidity, after 68-76 h, the living cell number is increased by 90-180% about, and the cell viability is increased back to 90% or above.

Description

technical field [0001] The present invention relates to the field of cell culture, in particular to a recovery method of CAR-T cells. Background technique [0002] CAR-T, the full name is Chimeric Antigen Receptor T-Cell Immunotherapy, chimeric antigen receptor T cell immunotherapy. This is a new type of cell therapy that has been around for many years, but has only been improved and used clinically in recent years. Similar to other immunotherapies, its basic principle is to use the patient's own immune cells to eliminate cancer cells. [0003] When a tumor is discovered clinically, it is generally in the middle and late stage. At this time, the tumor cells in the patient's body are dominant, which seriously damages the immune function of the body. In such a microenvironment of the immune system, the function of DC cells is impaired, and the efficiency of activating T cells is low. , the ability to attack cancer cells is not enough, and it is not precise enough; in additio...

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 Patents(China)
IPC IPC(8): C12N5/10
CPCC12N5/0636C12N2510/00
Inventor 曹路君丁兆
Owner SICHUAN HUIYU PHARMA