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Cryopreservation liquid and cryopreservation method for long-term preservation of stem cells and stem cell products

A technology for long-term preservation and cryopreservation, which is applied in the preservation, application, and animal husbandry of human or animal bodies, and can solve the problems of self-renewal and uncertain factors in the differentiation process of cultured stem cells, and achieves a method that is beneficial to long-term preservation and preparation Simple, effective in reducing the formation of ice crystals

Pending Publication Date: 2022-07-22
宁波建顺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the cryoprotectant is poorly selected, such as the use of animal-derived cryopreservation agents, it may bring uncertainties to the self-renewal and differentiation process of cultured stem cells

Method used

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  • Cryopreservation liquid and cryopreservation method for long-term preservation of stem cells and stem cell products
  • Cryopreservation liquid and cryopreservation method for long-term preservation of stem cells and stem cell products
  • Cryopreservation liquid and cryopreservation method for long-term preservation of stem cells and stem cell products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A cryopreservation solution for long-term preservation of stem cells and their products, including the following raw materials by weight: 10 parts of low-molecular-weight dextran, 2 parts of hydroxyethyl starch, 15 parts of chitosan, 3 parts of glycerol, and 3 parts of dimethyl sulfoxide 20 parts of glucose injection, 10 parts of compound amino acid injection and 20 parts of compound electrolyte injection.

[0029] The compound amino acid injection contains 6 g of L-leucine, 4 g of L-lysine acetate, 7 g of glycine, 3 g of methionine, 1 g of tyrosine, 2 g of glutamic acid, 2 g of threonine, and benzene per 1000 mL. 1 g of alanine, 1 g of aspartic acid, 1 g of cysteine, 1 g of serine, and 0.2 g of sodium bisulfite.

[0030] Compound electrolyte injection contains 3 g of calcium chloride dihydrate, 6 g of sodium chloride, 1 g of sodium gluconate, 1 g of sodium bicarbonate, 0.5 g of potassium chloride and 0.5 g of magnesium chloride per 1000 mL.

[0031] The glucose inject...

Embodiment 2

[0037] A cryopreservation solution for long-term preservation of stem cells and their products, including the following raw materials by weight: 15 parts of low molecular weight dextran, 8 parts of hydroxyethyl starch, 10 parts of chitosan, 6 parts of glycerol, 6 parts of dimethyl sulfoxide 30 parts of glucose injection, 15 parts of compound amino acid injection and 30 parts of compound electrolyte injection.

[0038] The compound amino acid injection contains 9 g of L-leucine, 8 g of L-lysine acetate, 9 g of glycine, 5 g of methionine, 3 g of tyrosine, 4 g of glutamic acid, 4 g of threonine, and benzene per 1000 mL. Alanine 3g, aspartic acid 3g, cysteine ​​3g, serine 3g and sodium bisulfite 0.4g.

[0039] Compound electrolyte injection contains 6 g of calcium chloride dihydrate, 9 g of sodium chloride, 3 g of sodium gluconate, 3 g of sodium bicarbonate, 0.8 g of potassium chloride and 0.8 g of magnesium chloride per 1000 mL.

[0040] The glucose injection contains 80 g of gl...

Embodiment 3

[0046] A cryopreservation solution for long-term preservation of stem cells and their products, including the following raw materials by weight: 12 parts of low molecular weight dextran, 6 parts of hydroxyethyl starch, 14 parts of chitosan, 4 parts of glycerol, 4 parts of dimethyl sulfoxide 16 parts of glucose injection, 13 parts of compound amino acid injection and 25 parts of compound electrolyte injection.

[0047] The compound amino acid injection contains 8 g of L-leucine, 5 g of L-lysine acetate, 8 g of glycine, 4 g of methionine, 2 g of tyrosine, 3 g of glutamic acid, 3 g of threonine, and benzene per 1000 mL. Alanine 2g, aspartic acid 2g, cysteine ​​2g, serine 2g and sodium bisulfite 0.3g.

[0048] Compound electrolyte injection contains 5 g of calcium chloride dihydrate, 8 g of sodium chloride, 2 g of sodium gluconate, 2 g of sodium bicarbonate, 0.6 g of potassium chloride and 0.6 g of magnesium chloride per 1000 mL.

[0049] The glucose injection contains 60 g of gl...

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Abstract

The invention discloses a cryopreservation solution and a cryopreservation method for long-term preservation of stem cells and stem cell products. The cryopreservation liquid is prepared from the following raw materials in parts by weight: 10 to 15 parts of low-molecular dextran, 2 to 8 parts of hydroxyethyl starch, 10 to 15 parts of chitosan, 3 to 6 parts of glycerol, 3 to 6 parts of dimethyl sulfoxide, 20 to 30 parts of glucose injection, 10 to 15 parts of compound amino acid injection and 20 to 30 parts of compound electrolyte injection. Under the combined action of the low-molecular dextran, the hydroxyethyl starch, the glucose injection, the compound amino acid injection and the compound electrolyte injection, the system balance of ion osmotic pressure and colloid osmotic pressure is maintained, an environmental condition beneficial to the survival of the stem cells is formed, and the long-term preservation of the stem cells is facilitated.

Description

technical field [0001] The invention relates to a cryopreservation solution and a cryopreservation method for long-term preservation of stem cells and their products, belonging to the technical field of cryopreservation solutions. Background technique [0002] Cryopreservation of cells refers to the preservation of cells in a low temperature environment to temporarily release the cells from the growth state and maintain their cell characteristics. Cells are preserved by freezing, which is less expensive, and can be resuscitated and cultured when needed. At the same time, cryopreservation can also avoid the loss of cell species due to cell contamination during culture. [0003] As a protective reagent that must be used in cell cryopreservation, the cell cryopreservation solution is used to suspend the cells that need to be cryopreserved and supply the nutrients needed for cell life metabolism. Cells are protected to prevent or reduce damage to cells caused by ice crystal fo...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/021
Inventor 张顺阮春燕施晓峰蔡挺张沛瑶
Owner 宁波建顺生物科技有限公司
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