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T lymphocyte cryopreservation solution, preparation method thereof and cell cryopreservation method

A technology of lymphocyte and cryopreservation method, applied in the field of cell cryopreservation, T lymphocyte cryopreservation solution and preparation thereof, can solve the problems of inability to meet the volume of cryopreservation solution, limited autologous plasma sampling, etc., and is helpful for rapid application. and promotion, meeting the needs of large-scale cryopreservation of cells, and the effect of simple operation

Inactive Publication Date: 2020-09-15
广州佑源生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used cryopreservation solution formula is prepared by dimethyl sulfoxide, autologous plasma and cell culture medium in a weight ratio of 1:4:5. Although this cryopreservation solution can maintain the proliferation rate of cells, However, due to limited autologous plasma materials, when a large number of cells are expanded and frozen, this formula cannot meet the volume of cryopreservation solution required for cell cryopreservation

Method used

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  • T lymphocyte cryopreservation solution, preparation method thereof and cell cryopreservation method
  • T lymphocyte cryopreservation solution, preparation method thereof and cell cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Weigh the following raw materials in parts by weight: 60 parts of amino acids, 350 parts of human serum albumin, 120 parts of inorganic salts, vitamin D 2 5 servings, Vitamin K 2 5 servings, Vitamin K 3 5 parts, 8 parts of docosahexaenoic acid, 8 parts of eicosapentaenoic acid, 15 parts of insulin, 150 parts of sodium pyruvate, and 2 parts of glutathione. Mix each raw material to make a serum substitute, called Get the following raw materials in parts by weight: 10 parts of dimethyl sulfoxide, 50 parts of serum-free cell culture medium, 40 parts of serum substitute, replace dimethyl sulfoxide, serum-free cell culture medium and gained serum The mixture was made into a cryopreservation solution.

[0034] In order to further illustrate the beneficial effects of the present invention, the present invention has carried out the experiment of following cell cryopreservation and resurrection:

[0035] (1) Prepare four freezing solutions:

[0036] ①The domestic cell cryopre...

Embodiment 2

[0046] Weigh the following raw materials in parts by weight: 50 parts of amino acids, 300 parts of human serum albumin, 80 parts of inorganic salts, vitamin D 2 3 servings, Vitamin K 2 3 servings, Vitamin K 3 3 parts, 3 parts of docosahexaenoic acid, 3 parts of eicosapentaenoic acid, 10 parts of insulin, 100 parts of sodium pyruvate, and 1 part of glutathione. Mix each raw material to make a serum substitute, called Get the following raw materials in parts by weight: 10 parts of dimethyl sulfoxide, 50 parts of serum-free cell culture medium, 40 parts of serum substitute, replace dimethyl sulfoxide, serum-free cell culture medium and gained serum The mixture was made into a cryopreservation solution.

Embodiment 3

[0048] Weigh the following raw materials in parts by weight: 55 parts of amino acids, 320 parts of human serum albumin, 100 parts of inorganic salts, vitamin D 2 4 servings, Vitamin K 2 4 servings, Vitamin K3 4 parts, 6 parts of docosahexaenoic acid, 6 parts of eicosapentaenoic acid, 12 parts of insulin, 120 parts of sodium pyruvate, and 1.5 parts of glutathione. Mix each raw material to make a serum substitute, called Get the following raw materials in parts by weight: 10 parts of dimethyl sulfoxide, 50 parts of serum-free cell culture medium, 40 parts of serum substitute, replace dimethyl sulfoxide, serum-free cell culture medium and gained serum The mixture was made into a cryopreservation solution.

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Abstract

The invention discloses a T lymphocyte cryopreservation solution and a preparation method and a cryopreservation method thereof. The T lymphocyte cryopreservation liquid comprises dimethyl sulfoxide,a serum-free cell culture medium and a serum substitute, and the serum substitute comprises the following components in parts by weight: amino acid, human serum albumin, inorganic salt, vitamin, docosahexaenoic acid, eicosapentaenoic acid, insulin, sodium pyruvate and glutathione. The T lymphocyte cryopreservation liquid provided by the invention is simple in components, simple in preparation process, low in cost and easy for industrial production; the cryopreservation liquid provided by the invention does not influence the characteristics of cells and the proliferation capacity of the cells,maintains the high recovery viability of the T lymphocytes, can better maintain the proliferation speed of the cells after cryopreservation recovery, can meet the requirements of large-scale cryopreservation of the cells and is beneficial to rapid application and popularization of a cell cryopreservation technology; the T lymphocyte cryopreservation method provided by the invention has the advantages of simple and rapid operation.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a T lymphocyte cryopreservation solution, a preparation method thereof and a cell cryopreservation method. Background technique [0002] T lymphocytes are derived from pluripotent stem cells of bone marrow. Mature T cells are distributed to the thymus-dependent areas of peripheral immune organs to settle in the peripheral immune organs through blood flow, and can be recirculated through lymphatic vessels, peripheral blood and tissue fluid to exert cellular immunity and immune regulation and other functions. At present, stem cell and immune cell technologies are widely used, and storage of stem cells and immune cells is an important link in the entire cell industry chain. The cryopreservation of T lymphocytes requires the use of protective agents to prevent the cells from being damaged by ice crystals, dehydration and water absorption during cryopreservation and recovery. A...

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 崔冬月刘向阳
Owner 广州佑源生物工程有限公司
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