Neural stem cell cryoprotectant and using method thereof

A neural stem cell and cryopreservation technology, applied in the field of neural stem cell cryopreservation, can solve the problems affecting the effect of neural stem cell cryopreservation and resuscitation, difficulty in determining the number of cells, and single composition of the cryopreservation solution. The effect of maintaining cell integrity and protecting cells

Active Publication Date: 2018-05-08
JILIN TUO HUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the intimate connection between neural stem cells and the combined effect of signals and diameters affect the cryopreservation and recovery effect of neural stem cells, and it is difficult to determine the number of cells, and the survival rate after recovery can only reach about 80%. For neural stem cells, the cryopreservation solution is composed of DMEMF12 with a volume fraction of 70%, serum with a volume fraction of 20%, and DMSO with a volume fraction of 10%.
Due to the relatively simple composition of the cryopreservation solution and the lack of necessary maintenance factors, the cell viability after recovery can only reach about 70%.

Method used

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  • Neural stem cell cryoprotectant and using method thereof
  • Neural stem cell cryoprotectant and using method thereof
  • Neural stem cell cryoprotectant and using method thereof

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Embodiment 1

[0039] The cryopreservation solution in the present embodiment consists of DMEM / F12 basal medium, dimethyl sulfoxide (DMSO), fetal calf serum, human serum albumin, Additives, epidermal growth factor and basic fibroblast growth factor were mixed and prepared, wherein: the addition amount of DMEM / F12 basal medium accounted for 50% of the volume percentage of the freezing solution; the addition amount of DMSO accounted for the volume percentage of the freezing solution was 10%; the added amount of fetal bovine serum accounted for 20% by volume of the frozen storage solution; the added amount of human albumin solution accounted for 18% by volume of the frozen storage solution; The amount of the additive solution added to the volume percentage of the frozen storage solution is 2%; the frozen storage solution contains epidermal growth factor with a concentration of 20ng / ml; the frozen storage solution contains a concentration of 20ng / ml basic fibroblast growth Factors (the volume o...

Embodiment 2

[0053] In the present embodiment, the cryopreservation solution consists of DMEM / F12 basal medium, DMSO, fetal bovine serum, human serum albumin, Additives, epidermal growth factor and basic fibroblast growth factor were mixed and prepared, wherein: the addition amount of DMEM / F12 basal medium accounted for 50% of the volume percentage of the freezing solution; the addition amount of DMSO accounted for the volume percentage of the freezing solution was 10%; the added amount of fetal bovine serum accounts for 18% of the volume percentage of the frozen storage solution; the added amount of human serum albumin accounts for 20% of the volume percentage of the frozen storage solution; The amount of additive added accounts for 2% of the volume percentage of the frozen storage solution; the frozen storage solution contains epidermal growth factor with a concentration of 20ng / ml; the frozen storage solution contains basic fibroblast growth factor with a concentration of 20ng / ml .

...

Embodiment 3

[0056] In the present embodiment, the cryopreservation solution consists of DMEM / F12 basal medium, DMSO, fetal bovine serum, human serum albumin, Additives, epidermal growth factor and basic fibroblast growth factor were mixed and prepared, wherein: the addition amount of DMEM / F12 basal medium accounted for 60% of the volume percentage of the freezing solution; the addition amount of DMSO accounted for the volume percentage of the freezing solution was 10%; the added amount of fetal bovine serum accounts for 15% of the volume percentage of the frozen storage solution; the added amount of human serum albumin accounts for 13% of the volume percentage of the frozen storage solution; The amount of additive added accounts for 2% of the volume percentage of the frozen storage solution; the frozen storage solution contains epidermal growth factor with a concentration of 20ng / ml; the frozen storage solution contains basic fibroblast growth factor with a concentration of 20ng / ml .

...

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Abstract

The invention discloses a neural stem cell cryoprotectant and a using method thereof. The cryoprotectant contains a DMEM/F12 basal culture medium, dimethyl sulfoxide, serum, albumin, a B27- additive,epidermal growth factors and alkaline fibroblast growth factors. The cryoprotectant is low in cost, and solves the problem that the anabiosis survival rate after cryopreservation of neural stem cellsis low; after the cells are cryopreservated for one year by adopting the neural stem cell cryoprotectant and the using method thereof disclosed by the invention, the cell viability still reaches up to95% higher, and the attribute and differentiative capacity of the neural stem cells are not influenced.

Description

technical field [0001] The invention belongs to the field of cell technology, and specifically discloses a neural stem cell cryopreservation solution and a method for using the same. Background technique [0002] Neural stem cell (Neural stem cell, NSC), as a kind of stem cell, has received more and more attention in the field of neuroscience in recent years, and the implementation of cell replacement therapy is a newly developed treatment strategy with broad application prospects. The discovery of neural stem cells and the success of in vitro culture make it possible to effectively treat central nervous system injuries and neurodegenerative diseases in the future. [0003] Functional loss caused by central nervous system injury such as cerebral infarction, cerebral hemorrhage, brain trauma, spinal cord injury, etc., and neurodegenerative diseases such as Parkinson's disease, amyotrophic lateral sclerosis, etc., is a difficult problem in the field of clinical treatment. Tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 毕薇薇李超姜丽君
Owner JILIN TUO HUA BIOTECH
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