Cell freezing medium for human neural stem cells

Abstract

The invention discloses a cell freezing medium for human neural stem cells, which is composed of a freezing medium A and a freezing medium B. The freezing medium A is composed of the following components: 10%-30% of albumin, 4.0-8.0 g / L of glucose, 0.4-0.8 mg / L of vitamin B12, 0.365 mg / L of glutamine, 6.0-9.0 g / L of sodium chloride and the like; the cryopreservation solution B is prepared from the following components: 10 to 20 percent of dimethyl sulfoxide, 4.0 to 8.0 g / L of glucose, 0.4 to 0.8 mg / L of vitamin B12 and the like. The invention also provides a method for cryopreserving human neural stem cells by using the cryopreservation liquid. According to the cell cryopreservation liquid and the cryopreservation method, serum is not contained, the pollution risk of animal-derived pathogenic microorganisms is avoided, the cryopreservation effect is excellent, the cell viability after resuscitation is high, and cell proliferation and cell functional characteristics are not influenced. The invention is of great significance to cryopreservation and research of human neural stem cells.

Description

technical field

[0001] The invention relates to a cell cryopreservation solution for human neural stem cells, belonging to the technical field of cell cryopreservation. Background technique

[0002] Cell cryopreservation is an important link in cell culture technology, and it is an essential link in the construction of cell banks. Cell cryopreservation solution is a kind of protective solution that must be used when cells are frozen. Its purpose is to fully mix the cells that need to be frozen with the protective solution into a cell suspension, and prevent or reduce the freezing process during the rapid cooling and freezing of cells. damage to cells by ice crystals. At the same time, during the long-term cryogenic storage of cells, it can supply the nutrients needed for cell life metabolism.

[0003] The currently commonly used cryopreservation solution is generally a mixture of culture medium, serum and dimethyl sulfoxide (DMSO) in a certain proportion. The disadvantage ...

Examples

Embodiment 1

[0030] Example 1 Preparation of cell cryopreservation solution for human neural stem cells

[0031] It is composed of cryopreservation solution A and cryopreservation solution B, wherein 1L of cryopreservation solution A is composed of the following components: 20% albumin, 6.0g / L glucose, 0.6mg / L vitamin B12, 0.365mg / L Glutamine, 7.5g / L sodium chloride, 0.025g / L magnesium chloride, 0.3g / L potassium chloride, and the balance is water.

[0032] 1L of freezing solution B is composed of the following components: 15% dimethyl sulfoxide, 6.0g / L glucose, 0.6mg / L vitamin B12, 0.365mg / L glutamine, 7.5g / L sodium chloride , 0.025g / L magnesium chloride, 0.3g / L potassium chloride, and the balance is water.

[0033] The preparation method of cell cryopreservation solution is as follows: prepare cryopreservation solution A and cryopreservation solution B respectively; add each component into ultrapure water, mix and dissolve, and shake and mix with an oscillator for 2 to 3 minutes during t...

Embodiment 2

[0034] Example 2 Preparation of cell cryopreservation solution for human neural stem cells

[0035] It consists of cryopreservation solution A and cryopreservation solution B, wherein, cryopreservation solution A is composed of the following components: 10% albumin, 8.0g / L glucose, 0.4mg / L vitamin B12, 0.365mg / L glutamine Amide, 9.0g / L sodium chloride, 0.015g / L magnesium chloride, 0.4g / L potassium chloride, and the balance is water.

[0036] Freezing solution B is composed of the following components: 10% dimethyl sulfoxide, 8.0g / L glucose, 0.4mg / L vitamin B12, 0.365mg / L glutamine, 9.0g / L sodium chloride, 0.015 g / L magnesium chloride, 0.4g / L potassium chloride, and the balance is water.

[0037] The preparation method of cell cryopreservation solution is as follows: prepare cryopreservation solution A and cryopreservation solution B respectively; add each component into ultrapure water, mix and dissolve, and shake and mix with an oscillator for 2 to 3 minutes during the disso...

Embodiment 3

[0038] Example 3 Preparation of cell cryopreservation solution for human neural stem cells

[0039] It is composed of cryopreservation solution A and cryopreservation solution B, wherein, cryopreservation solution A is composed of the following components: 30% albumin, 4.0g / L glucose, 0.8mg / L vitamin B12, 0.365mg / L glutamine Amide, 6.0g / L sodium chloride, 0.030g / L magnesium chloride, 0.2g / L potassium chloride, and the balance is water.

[0040] Freezing solution B is composed of the following components: 20% dimethyl sulfoxide, 4.0g / L glucose, 0.8mg / L vitamin B12, 0.365mg / L glutamine, 6.0g / L sodium chloride, 0.030 g / L magnesium chloride, 0.2g / L potassium chloride, and the balance is water.

[0041] The preparation method of cell cryopreservation solution is as follows: prepare cryopreservation solution A and cryopreservation solution B respectively; add each component into ultrapure water, mix and dissolve, and shake and mix with an oscillator for 2 to 3 minutes during the diss...

technical field

[0001] The invention relates to a cell cryopreservation solution for human neural stem cells, belonging to the technical field of cell cryopreservation. Background technique

[0002] Cell cryopreservation is an important link in cell culture technology, and it is an essential link in the construction of cell banks. Cell cryopreservation solution is a kind of protective solution that must be used when cells are frozen. Its purpose is to fully mix the cells that need to be frozen with the protective solution into a cell suspension, and prevent or reduce the freezing process during the rapid cooling and freezing of cells. damage to cells by ice crystals. At the same time, during the long-term cryogenic storage of cells, it can supply the nutrients needed for cell life metabolism.

[0003] The currently commonly used cryopreservation solution is generally a mixture of culture medium, serum and dimethyl sulfoxide (DMSO) in a certain proportion. The disadvantage ...

Examples

Embodiment 1

[0030] Example 1 Preparation of cell cryopreservation solution for human neural stem cells

[0031] It is composed of cryopreservation solution A and cryopreservation solution B, wherein 1L of cryopreservation solution A is composed of the following components: 20% albumin, 6.0g / L glucose, 0.6mg / L vitamin B12, 0.365mg / L Glutamine, 7.5g / L sodium chloride, 0.025g / L magnesium chloride, 0.3g / L potassium chloride, and the balance is water.

[0032] 1L of freezing solution B is composed of the following components: 15% dimethyl sulfoxide, 6.0g / L glucose, 0.6mg / L vitamin B12, 0.365mg / L glutamine, 7.5g / L sodium chloride , 0.025g / L magnesium chloride, 0.3g / L potassium chloride, and the balance is water.

[0033] The preparation method of cell cryopreservation solution is as follows: prepare cryopreservation solution A and cryopreservation solution B respectively; add each component into ultrapure water, mix and dissolve, and shake and mix with an oscillator for 2 to 3 minutes during t...

Embodiment 2

[0034] Example 2 Preparation of cell cryopreservation solution for human neural stem cells

[0035] It consists of cryopreservation solution A and cryopreservation solution B, wherein, cryopreservation solution A is composed of the following components: 10% albumin, 8.0g / L glucose, 0.4mg / L vitamin B12, 0.365mg / L glutamine Amide, 9.0g / L sodium chloride, 0.015g / L magnesium chloride, 0.4g / L potassium chloride, and the balance is water.

[0036] Freezing solution B is composed of the following components: 10% dimethyl sulfoxide, 8.0g / L glucose, 0.4mg / L vitamin B12, 0.365mg / L glutamine, 9.0g / L sodium chloride, 0.015 g / L magnesium chloride, 0.4g / L potassium chloride, and the balance is water.

[0037] The preparation method of cell cryopreservation solution is as follows: prepare cryopreservation solution A and cryopreservation solution B respectively; add each component into ultrapure water, mix and dissolve, and shake and mix with an oscillator for 2 to 3 minutes during the disso...

Embodiment 3

[0038] Example 3 Preparation of cell cryopreservation solution for human neural stem cells

[0039] It is composed of cryopreservation solution A and cryopreservation solution B, wherein, cryopreservation solution A is composed of the following components: 30% albumin, 4.0g / L glucose, 0.8mg / L vitamin B12, 0.365mg / L glutamine Amide, 6.0g / L sodium chloride, 0.030g / L magnesium chloride, 0.2g / L potassium chloride, and the balance is water.

[0040] Freezing solution B is composed of the following components: 20% dimethyl sulfoxide, 4.0g / L glucose, 0.8mg / L vitamin B12, 0.365mg / L glutamine, 6.0g / L sodium chloride, 0.030 g / L magnesium chloride, 0.2g / L potassium chloride, and the balance is water.

[0041] The preparation method of cell cryopreservation solution is as follows: prepare cryopreservation solution A and cryopreservation solution B respectively; add each component into ultrapure water, mix and dissolve, and shake and mix with an oscillator for 2 to 3 minutes during the diss...