Method for preserving immune cells and cell cryopreservation solution

A technology of immune cells and storage methods, applied in the direction of preservation, application, animal husbandry, etc. of human or animal bodies

Inactive Publication Date: 2018-09-11
广州沙艾生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The immune ability of the human immune cells used in immunotherapy is closely related to the health of the human body. If the immune cells with good health can be preserved, they can be better used in anti-tumor therapy and help to achieve better therapeutic effects ; There are still few effective preservation methods and cryopreservation solutions for the effective preservation of immune cells, and there is an urgent need for effective immune cell preservation methods and cell cryopreservation solutions to preserve immune cells

Method used

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  • Method for preserving immune cells and cell cryopreservation solution
  • Method for preserving immune cells and cell cryopreservation solution
  • Method for preserving immune cells and cell cryopreservation solution

Examples

Experimental program
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Effect test

Embodiment 1

[0033] A cryopreservation solution of immune cells is calculated by volume percentage and contains the following components: 41% 1640 medium, 22% fetal bovine serum and 8% dimethyl sulfoxide.

[0034] A storage method of immune cells includes the following steps:

[0035] S1. Extract fresh peripheral blood (PBMC) by routine aseptic operation, and dilute the above-mentioned peripheral blood with 1640 medium and shake it evenly to form a diluent, then 5-9ml of diluent and 15ml of lymphocyte separation liquid Mix, the upper layer is blood thinner, and the lower layer is lymphocyte separation liquid;

[0036] S2. Then centrifuge the above mixture (2000rpm, 20min, 22°C, and adjust the acceleration to 0) to form a stratification. After the centrifugation is completed, absorb the mononuclear cells in the albuginea layer, and then add an appropriate amount of 1640 Medium, and beat well;

[0037] S3. Centrifuge the above solution (1500rpm, 10min, 4°C, and adjust the vertical acceleration to 9...

Embodiment 2

[0043] A cryopreservation solution of immune cells is calculated by volume percentage and contains the following components: 75% 1640 medium, 40% fetal bovine serum and 17% dimethyl sulfoxide.

[0044] A storage method of immune cells includes the following steps:

[0045] S1. Extract fresh peripheral blood (PBMC) by routine aseptic operations, and dilute the above-mentioned peripheral blood with 1640 medium and shake it to form a dilution, then mix 9ml of the dilution with 15ml of the lymphocyte separation solution, The upper layer is blood thinner, and the lower layer is lymphocyte separation liquid;

[0046] S2. Then centrifuge the above-mentioned mixture (2000rpm, 21min, 22°C, and adjust the vertical acceleration to 0) to form a stratification. After centrifugation is completed, absorb the mononuclear cells in the albuginea layer, and then add an appropriate amount of 1640 Medium, and beat well;

[0047] S3. Centrifuge the above solution (1500rpm, 8min, 4°C, and adjust the vertic...

Embodiment 3

[0053] A cryopreservation liquid for immune cells, in terms of volume percentage, contains the following components: 56% 1640 medium, 32% fetal bovine serum and 12% dimethyl sulfoxide.

[0054] A storage method of immune cells includes the following steps:

[0055] S1. Extract fresh peripheral blood (PBMC) by routine aseptic operations, and dilute the above-mentioned peripheral blood with 1640 medium and shake it evenly to form a dilution, and then mix 7ml of the dilution with 15ml of the lymphocyte separation solution, The upper layer is blood thinner, and the lower layer is lymphocyte separation liquid;

[0056] S2. Then centrifuge the above-mentioned mixture (2000rpm, 22min, 22°C, and adjust the vertical acceleration to 0) to form a stratification. After the centrifugation is completed, absorb the mononuclear cells in the albuginea layer, and then add an appropriate amount of 1640 Medium, and beat well;

[0057] S3. Centrifuge the above solution (1500rpm, 12min, 4°C, and adjust th...

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Abstract

The invention relates to the technical field of cell preservation, in particular to a method for preserving immune cells and a cell cryopreservation solution. The method comprises the following stepsof firstly, collecting peripheral blood; secondly, separating the peripheral blood; thirdly, adding the cell cryopreservation solution into the peripheral blood for cryopreservation. The cell cryopreservation solution contains, by volume, a cell basal culture medium, fetal bovine serum and dimethyl sulfoxide. The method for preserving the immune cells is simple in step and easy to operate, the fetal bovine serum has the advantages of wide source and low price, the cost decrease is facilitated, and the method is convenient to popularize and apply; the immune cells are cryopreserved and recovered in the cell cryopreservation solution and then subjected to induced culture, and the survival rate is 90% or above, it is shown through the detection on various indexes of the immune cells that thefunctions of the immune cells keep complete, and it shows that the cell cryopreservation solution can maintain the physiological functions of the immune cells well and has great significance for clinical treatment.

Description

Technical field [0001] The invention relates to the technical field of cell culture, in particular to a storage method of immune cells and a cell cryopreservation solution. Background technique [0002] In recent years, due to the increasingly serious environmental air pollution, the intensification of social competition and the increasing pressure of people’s lives, people’s bodies and minds have been in a state of tension for a long time, which has seriously affected people’s health. Moreover, most wage earners have been lacking for a long time. Exercise has led to a continuous decline in physical fitness, and the body's resistance is gradually weakened; more and more people have sub-health problems in their bodies, and more and more people have various problems such as endocrine disorders and decreased resistance; moreover, due to the current food The illegal addition and abuse of additives in cosmetics, heavy metal content and pesticide residues are still quite serious, all o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 徐智峰张新
Owner 广州沙艾生物科技有限公司
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