Immune cell cryopreservation method

A technology of immune cell and cryopreservation method, applied in the field of immune cell cryopreservation, can solve problems such as unfavorable patient health, allergic reaction, immune rejection, etc., and achieve the effect of reducing possibility, promoting proliferation ability, and reducing usage

Inactive Publication Date: 2021-05-07
浙江卫未生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most of the existing cell cryopreservation solutions are mixed with dimethyl sulfoxide and animal serum. Due to the high content of dimethyl sulfoxide, it is quite toxic, which is not conducive to the health of patients, and animal serum contains a large amount of foreign substances. protein, risk of infection, allergic reaction or immune rejection

Method used

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  • Immune cell cryopreservation method
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  • Immune cell cryopreservation method

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A method for cryopreserving immune cells, comprising the steps of:

[0036] S1. Obtain immune cells

[0037] Mix normal saline, Ficoll separation solution with peripheral blood or umbilical cord blood, centrifuge at 2000rpm for 8 minutes at a temperature of 37°C, absorb the middle buffy coat, wash with normal saline twice, and separate to obtain immune cells;

[0038] S2. Obtain human autologous plasma

[0039] Take human peripheral blood or umbilical cord blood and place it in a sterile centrifuge tube, centrifuge at 15°C and 2000rpm for 4 minutes, take the upper layer of plasma and inactivate it at 62°C, transfer it to a new centrifuge tube, and centrifuge at 3000rpm , centrifuged for 15 minutes, and the supernatant was drawn to obtain human autologous plasma;

[0040] S3. Prepare cell cryopreservation solution

[0041] The cell freezing solution includes dimethyl sulfoxide, cell culture medium, human serum albumin, carbohydrates, resveratrol, hydroxyethyl starch, ...

Embodiment 2

[0049] A method for cryopreserving immune cells, comprising the steps of:

[0050] S1. Obtain immune cells

[0051] Mix normal saline, Ficoll separation solution with peripheral blood or umbilical cord blood, centrifuge at 2000rpm for 10 minutes at 30°C, absorb the middle buffy coat, wash with normal saline twice, and separate to obtain immune cells;

[0052] S2. Obtain human autologous plasma

[0053]Take human peripheral blood or umbilical cord blood and place it in a sterile centrifuge tube, centrifuge at 20°C and 2000rpm for 6 minutes, take the upper layer of plasma and inactivate it at 65°C, transfer it to a new centrifuge tube, and centrifuge at 5000rpm , centrifuged for 20 minutes, and the supernatant was drawn to obtain human autologous plasma;

[0054] S3. Prepare cell cryopreservation solution

[0055] The cell freezing solution includes dimethyl sulfoxide, cell culture medium, human serum albumin, carbohydrates, resveratrol, hydroxyethyl starch, propylene glycol,...

Embodiment 3

[0063] A method for cryopreserving immune cells, comprising the steps of:

[0064] S1. Obtain immune cells

[0065] Mix normal saline, Ficoll separation solution with peripheral blood or umbilical cord blood, centrifuge at 2000rpm for 10 minutes at a temperature of 35°C, absorb the middle buffy coat, wash twice with normal saline, and separate to obtain immune cells;

[0066] S2. Obtain human autologous plasma

[0067] Take human peripheral blood or umbilical cord blood and place it in a sterile centrifuge tube, centrifuge at 20°C and 2000rpm for 4 minutes, take the upper layer of plasma and inactivate it at 64°C, transfer it to a new centrifuge tube, and centrifuge at 4000rpm , centrifuged for 17 minutes, and the supernatant was drawn to obtain human autologous plasma;

[0068] S3. Prepare cell cryopreservation solution

[0069] The cell freezing solution includes dimethyl sulfoxide, cell culture medium, human serum albumin, carbohydrates, resveratrol, hydroxyethyl starch,...

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Abstract

The invention provides an immune cell cryopreservation method. The immune cell cryopreservation method comprises the following steps: S1, acquiring immune cells; S2, acquiring human autologous plasma; S3, preparing a cell freezing medium; S4, carrying out cell cryopreservation. According to the immune cell cryopreservation method provided by the invention, the immune cells can be effectively prevented from being damaged by freezing, the recovery survival rate of the immune cells is improved, and the resveratrol added into the cell cryopreservation liquid can promote the multiplication capacity of the recovered immune cells, so the activity of the recovered cryopreserved immune cells is improved; hydroxyethyl starch, propylene glycol and polyethylene glycol can replace the protection effect of dimethyl sulfoxide, the use amount of dimethyl sulfoxide is reduced, it is guaranteed that when cells are close to the freezing point, water in the cells cannot crystallize, damage to the cells is small, and foreign protein cannot be introduced into human autologous plasma added into the cell freezing medium, so the possibility of animal disease source pollution is reduced, and the potential safety hazard of exogenous serum is avoided.

Description

technical field [0001] The invention relates to a cryopreservation method for immune cells, which belongs to the technical field of cell engineering. Background technique [0002] Immune cell therapy is a new type of autoimmune anti-cancer treatment. It uses biotechnology and biological agents to culture and amplify immune cells collected from patients in vitro and then reinfuse them back into patients to stimulate and enhance immunity. The body's own immune function, so as to achieve the purpose of treating tumors. For example, immune cells are frozen and resuscitated and reinfused immediately when needed. [0003] When cells are cryopreserved, osmotic protectants are often used to reduce the damage to the cells during the cryopreservation process. At present, the commonly used cryopreservation agent is dimethyl sulfoxide (DMSO), which can quickly penetrate into the cell interior and improve the cell membrane. The permeability to water allows water to seep out of the cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 陈锦阳林志伟刘军权史军
Owner 浙江卫未生物医药科技有限公司
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