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Immunization cell frozen stock solution, preparation method and application

A technology of immune cells and cryopreservation solution, which is applied in applications, preservation of human or animal bodies, animal husbandry, etc. It can solve the problems of increasing animal pathogen pollution, long storage time of cells, and the impact of human adoptive immunotherapy, so as to achieve the protection of cryopreservation Effects of injury, reduction of likelihood, preservation of physiological function and biological properties

Active Publication Date: 2014-09-10
HANGZHOU S EVANS BIOSCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, domestic cryopreservation methods mostly use DMSO, animal serum and cell culture medium in different proportions. At the same time, it increases the possibility of animal pathogen contamination, which will have a certain impact on human adoptive immunotherapy

Method used

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  • Immunization cell frozen stock solution, preparation method and application
  • Immunization cell frozen stock solution, preparation method and application
  • Immunization cell frozen stock solution, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Preparation of Human Donor Plasma

[0036] (1) Blood draw: Select two volunteers (A and B), draw 50ml of peripheral blood with 50ml syringe respectively, and anticoagulate with heparin (other anticoagulation methods such as sodium citrate or EDTA can also be used).

[0037] (2) A small amount of peripheral blood was taken for specific virus tests such as Treponema pallidum, cytomegalovirus, hepatitis B virus, hepatitis C virus, HIV, and Epstein-Barr virus, all of which were negative (Table 1), proving that both volunteers were Healthy individuals who do not carry the above-mentioned germs or viruses.

[0038] Table 1 Volunteer blood test results

[0039] project

[0040] (3) Preparation of donor plasma: transfer 50ml of anticoagulated peripheral blood to two 50ml centrifuge tubes equally, centrifuge at 2500rpm for 15min; absorb the upper layer of plasma, put it in a water bath at 56°C for 30min, then centrifuge at 2500rpm for 5min, take The super...

Embodiment 2

[0041]Example 2: Preparation of immune cell cryopreservation solution 1 (fetal bovine serum + DMSO + RPMI-1640 medium)

[0042] According to volume percentage, 10% fetal bovine serum was mixed with 10% DMSO and 80% RPMI-1640 medium to prepare immune cell cryopreservation solution 1, which was stored at 4°C for later use.

Embodiment 3

[0043] Example 3: Preparation of Immune Cell Cryopreservation Solution 2 (Donor Plasma+DMSO+RPMI-1640 Medium)

[0044] According to volume percentage, 10% donor plasma was mixed with 10% DMSO and 80% RPMI-1640 medium to prepare immune cell cryopreservation solution 2, which was stored at 4°C for later use.

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PUM

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Abstract

The invention discloses an immunization cell frozen stock solution, a preparation method and an application, the immunization cell frozen stock solution contains dimethyl sulfoxide, human donor plasma and an immunization cell basic medium. The immunization cell frozen stock solution comprises the following components according to volume percentage: 5-10% of dimethyl sulfoxide, 10-90% of human donor plasma and 0.01-80% of immunization cell basic medium. According to the immunization cell frozen stock solution, no animal serum is contained, exogenous protein is not introduced, so that possibility of animal disease cause pollution is reduced; refrigeration damage of the immunization cell can be effectively avoided, physiology function and stability of biology characteristic after recovering the immunization cell can be kept, the immunization cell frozen stock solution can be directly used for clinical treatment, and is suitable for long-term refrigeration storage of human mononuclear cell and other various immunization cells.

Description

technical field [0001] The invention relates to the technical field of cell preservation, in particular to an immune cell cryopreservation solution, its preparation method and application. Background technique [0002] Cryopreservation of donor immune cells can maintain their vitality and function, which is of great significance for both clinical and basic research, especially when immune cells are used for retrospective research. Cryopreservation of immune cells can not only solve the problem of long induction time of existing immune cells and the need for multiple inductions and multiple blood collections from patients, but also preserve the immune cells in the best state of health and the strongest combat effectiveness for tumor treatment and anti-aging health care treatments. [0003] The main factors affecting the cryopreservation and resuscitation activity of immune cells include the method of freezing and resuscitation and the choice of cryogen. Cell damage is cause...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 项春生王大奎张翠翠
Owner HANGZHOU S EVANS BIOSCI LTD
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