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Cell cryopreservation solution and cryopreservation method

A cryopreservation method and technology of cryopreservation solution, which are applied in the preservation, application, and animal husbandry of human or animal bodies, can solve the problems of lack of strict control of technology, great influence on cell survival, and denaturation of intracellular proteins, etc. Anti-freeze damage ability, reduce the number of cell death, prevent the effect of cell loss

Inactive Publication Date: 2020-06-30
GUANGDONG PANGUARD CELL BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dimethyl sulfoxide (DMSO) is the most commonly used cryoprotectant in cell cryopreservation, but too high a concentration of DMSO has certain toxicity to cells and can cause intracellular protein denaturation, so the selection of an unsuitable DMSO concentration will have a negative effect on cells. The preservation of
The cells frozen in the existing cryopreservation solution have a low yield of viable cells after recovery, and the cryopreservation solution will cause great damage to the cells, which will directly lead to cell death
The method of cooling during cryopreservation has a great impact on cell survival, and the existing technology is not strictly controlled, resulting in an increase in cell death rate during the cooling process

Method used

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  • Cell cryopreservation solution and cryopreservation method
  • Cell cryopreservation solution and cryopreservation method
  • Cell cryopreservation solution and cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A cell cryopreservation solution, comprising the following raw materials:

[0045]

[0046] The balance is sodium chloride injection.

[0047] Wherein, the mass concentration of the sodium chloride injection is 0.9%.

[0048] A cell cryopreservation method, comprising the steps of:

[0049] (1) Get the cells in the exponential growth phase and centrifuge, discard the supernatant, the cells in the exponential growth phase are PBMC or stem cells or T lymphocytes;

[0050] (2) Resuspend with pre-frozen culture medium, and then centrifuge;

[0051] (3) Repeat step (2) twice, then resuspend with pre-frozen medium to adjust cell density;

[0052] (4) At 37°C, 5% CO 2 , incubate for 45 minutes under saturated humidity conditions;

[0053] (5) Centrifuge and discard the supernatant;

[0054] (6) Resuspend with cell freezing solution, then centrifuge, and discard the supernatant;

[0055] (7) Resuspend with cell freezing solution, adjust cell density, and transfer to c...

Embodiment 2

[0078] A cell cryopreservation solution, comprising the following raw materials:

[0079]

[0080]

[0081] The balance is sodium chloride injection.

[0082] Wherein, the mass concentration of the sodium chloride injection is 0.9%.

[0083] A cell cryopreservation method, comprising the steps of:

[0084] (1) Get the cells in the exponential growth phase and centrifuge, discard the supernatant, the cells in the exponential growth phase are PBMC or stem cells or T lymphocytes;

[0085] (2) Resuspend with pre-frozen culture medium, and then centrifuge;

[0086] (3) Repeat step (2) once, then resuspend with pre-frozen medium to adjust the cell density;

[0087] (4) At 37°C, 5% CO 2 , incubate for 30 min under saturated humidity conditions;

[0088] (5) Centrifuge and discard the supernatant;

[0089] (6) Resuspend with cell freezing solution, then centrifuge, and discard the supernatant;

[0090] (7) Resuspend with cell freezing solution, adjust cell density, and tr...

Embodiment 3

[0110] A cell cryopreservation solution, comprising the following raw materials:

[0111]

[0112] The balance is sodium chloride injection.

[0113] Wherein, the mass concentration of the sodium chloride injection is 0.9%.

[0114] A cell cryopreservation method, comprising the steps of:

[0115] (1) Get the cells in the exponential growth phase and centrifuge, discard the supernatant, the cells in the exponential growth phase are PBMC or stem cells or T lymphocytes;

[0116] (2) Resuspend with pre-frozen culture medium, and then centrifuge;

[0117] (3) Repeat step (2) twice, then resuspend with pre-frozen medium to adjust cell density;

[0118] (4) At 37°C, 5% CO 2 , incubate for 60 min under saturated humidity conditions;

[0119] (5) Centrifuge and discard the supernatant;

[0120] (6) Resuspend with cell freezing solution, then centrifuge, and discard the supernatant;

[0121] (7) Resuspend with cell freezing solution, adjust cell density, and transfer to cryop...

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Abstract

The invention relates to the technical field of cell cryopreservation. The invention relates to a cell cryopreservation solution and a cryopreservation method. Wherein the cell cryopreservation liquidis prepared from methyl cellulose, maltose, DMSO, human serum albumin, dextran, L-serine, i-inositol and sodium chloride injection, and in the cell cryopreservation solution, DMSO can prevent damagecaused by intracellular fluid ice crystal formation, osmotic pressure change, cell structure disorder and the like; methyl cellulose is used as a stabilizer and a tackifier, so that the damage to cells caused by ice crystals formed by free water in the cryopreservation process is reduced; maltose has good crystallization resistance and prevents damage of ice crystals to cells. Through the combination of the raw materials, the loss of cells caused by ice crystals can be prevented to the maximum extent, and the content of DMSO is reduced as much as possible, so that the cell cryopreservation solution has a relatively good protection capability on cells such as PBMC or stem cells or T lymphocytes, and is high in survival rate.

Description

technical field [0001] The invention relates to the technical field of cell cryopreservation, in particular to a cell cryopreservation solution and a cryopreservation method. Background technique [0002] At present, in the field of cell cryopreservation, in order to protect cells from freezing damage, it is widely used to add a cryoprotectant (CPA) to the cryopreservation solution. Dimethyl sulfoxide (DMSO) is the most commonly used cryoprotectant in cell cryopreservation, but too high a concentration of DMSO has certain toxicity to cells and can cause intracellular protein denaturation, so the selection of an unsuitable DMSO concentration will have a negative effect on cells. preservation has a great influence. The cells frozen in the existing cryopreservation solution have a low yield of viable cells after thawing, and the cryopreservation solution does a lot of damage to the cells, which will directly lead to cell death. The method of cooling during cryopreservation ha...

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 彭一洪李相鲁李莉何安涛
Owner GUANGDONG PANGUARD CELL BIOLOGICAL TECH CO LTD
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