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Use of epicatechin, cell cryopreservation solution and cell cryopreservation method

A technology of cryopreservation liquid and epicatechin, which can be used in the preservation, application, animal husbandry and other directions of human or animal body, and can solve the problems of factors such as lack of cell activity.

Active Publication Date: 2022-02-01
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the number of cells in this method is controllable, the cells are exposed to the freezing environment and lack factors to maintain cell viability, so the cell recovery rate is only about 70%.

Method used

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  • Use of epicatechin, cell cryopreservation solution and cell cryopreservation method
  • Use of epicatechin, cell cryopreservation solution and cell cryopreservation method
  • Use of epicatechin, cell cryopreservation solution and cell cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Prepare a complete medium for neural stem cells, which contains 88vol% of DMEM / F12 medium, 1vol% of N2 supplement, 1vol% of streptomycin-penicillin mixed solution, 20ng / ml of epidermal growth factor EGF and 20ng / ml of alkali fibroblast growth factor bFGF. The concentration of streptomycin in the streptomycin-penicillin mixed solution was 10,000mcg / ml, and the concentration of penicillin was 12,000Units / ml. Add 5 vol% dimethyl sulfoxide and 130 μg / ml epicatechin to the complete neural stem cell medium to obtain a cell cryopreservation solution. See Table 1 for details.

Embodiment 2～4 and comparative example 1～8

[0085] Except for the concentration of dimethyl sulfoxide and the concentration of epicatechin, other conditions are the same as in Example 1. See Table 1 for details.

experiment example

[0090] Experimental example-neural stem cell cryopreservation experiment

[0091] 1. Experimental method

[0092] Neural stem cells were self-extracted from the striatum of 12.5-day-old ICR mouse embryos, and then passaged and purified. When the neural stem cells entered the exponential growth phase, the cell spheres were collected, pipetted to separate the cell spheres into single cells, centrifuged, and the medium was discarded.

[0093] For cell counting, take 1×10 5 cell viability assay. The remaining part in 1×10 6 The concentration of each cell / ml was divided into cell cryopreservation tubes containing the cell cryopreservation solutions of Examples 1-4 and Comparative Examples 1-18 respectively.

[0094] The cells in the cell cryopreservation tube were loaded into the programmed cryopreservation box, kept overnight at -80°C, and transferred to a liquid nitrogen tank for cryopreservation for more than 1 day the next day.

[0095] Take 1×10 5 Cells were centrifuge...

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Abstract

The invention discloses an application of epicatechin, a cell cryopreservation solution and a cell cryopreservation method. Adding epicatechin as a component of the cell cryopreservation solution to the cell cryopreservation solution can improve the recovery rate of neural stem cells after cryopreservation.

Description

technical field [0001] The invention relates to the use of epicatechin, a cell cryopreservation solution and a cell cryopreservation method. Background technique [0002] For neurodegenerative diseases that cannot be cured by drugs such as Parkinson's, neural stem cell therapy has brought unprecedented light. Neural stem cells have unlimited potential for self-renewal and differentiation into neurons and glial cells. In the process of cell therapy, on the one hand, the neural stem cells in the patient's brain can be internally stimulated to differentiate and migrate to the diseased area, so as to restore the restricted physiological process; on the other hand, by transplanting functional neural stem cells in vitro, Replenishes cells that are missing or have lost normal function in the body. It has been widely recognized that transplanted cells are derived from induced neural stem cells derived from directed differentiation of pluripotent stem cells. However, so far, the n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0215A01N1/021
Inventor 贺喜白乙那木拉张静怡艾丽亚
Owner INNER MONGOLIA UNIVERSITY