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Mild enzyme cell digestive juice for stem cells

A digestive juice and stem cell technology, applied in the field of stem cell mild enzyme digestive juice, can solve the problems of donor contamination, complex components, etc., and achieve the effect of stable quality, high cell viability, and guaranteed batch-to-batch difference

Inactive Publication Date: 2019-03-08
YOCON BIOLOGY TECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since trypsin is derived from living organisms and its components are complex, the donor is easily contaminated by pathogens. Once this pollution is not completely removed and is brought into the production of trypsin, it may lead to serious consequences.

Method used

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  • Mild enzyme cell digestive juice for stem cells
  • Mild enzyme cell digestive juice for stem cells
  • Mild enzyme cell digestive juice for stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of mild enzyme cell digestion solution for stem cells

[0021] It is composed of the following concentration components, each concentration unit is mg / L: dipotassium hydrogen phosphate 60, potassium chloride 400, sodium bicarbonate 350, sodium chloride 8000, disodium hydrogen phosphate heptahydrate 90, glucose 1500, EDTA-4Na 350, 30 mild digestive enzymes for stem cells, the balance is water.

[0022] The preparation method is: take the above-mentioned components except water, classify and dissolve according to their respective dissolution characteristics, then mix, add water to make the final concentration of each component as described above, adjust the pH value to 7.2-7.4, that is, filter by industrial filter. At the same time, nitrogen protection is used for sub-packaging (to avoid oxidation of unstable components).

Embodiment 2

[0023] Example 2 Preparation of stem cell mild digestion enzyme cell digestion solution

[0024] It is composed of the following concentration components, each concentration unit is mg / L: dipotassium hydrogen phosphate 50, potassium chloride 450, sodium bicarbonate 300, sodium chloride 9000, disodium hydrogen phosphate heptahydrate 80, glucose 2000, EDTA-4Na 500, mild digestion enzyme for stem cells 10, the balance is water.

[0025] The preparation method is: take the above-mentioned components except water, classify and dissolve according to their respective dissolution characteristics, then mix, add water to make the final concentration of each component as described above, adjust the pH value to 7.2-7.4, that is, filter by industrial filter. At the same time, nitrogen protection is used for sub-packaging (to avoid oxidation of unstable components).

Embodiment 3

[0026] Example 3 Preparation of mild stem cell digestion enzyme cell digestion solution

[0027] It is composed of the following concentration components, each concentration unit is mg / L: dipotassium hydrogen phosphate 70, potassium chloride 350, sodium bicarbonate 400, sodium chloride 7000, disodium hydrogen phosphate heptahydrate 100, glucose 2000, EDTA-4Na 200, mild digestion enzyme for stem cells 50, the balance is water.

[0028] The preparation method is: take the above-mentioned components except water, classify and dissolve according to their respective dissolution characteristics, then mix, add water to make the final concentration of each component as described above, adjust the pH value to 7.2-7.4, that is, filter by industrial filter. , And at the same time nitrogen protection for sub-packaging (to avoid oxidation of unstable components).

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Abstract

The invention discloses a mild cell digestive enzyme cell digestive juice for stem cells. The digestive juice is composed of components at the following concentrations, according to a concentration unit mg / L: dipotassium hydrogen phosphate at 50 to 70, potassium chloride at 350 to 450, sodium hydrogen carbonate at 300 to 400, sodium chloride at 7,000 to 9,000, sodium hydrogen phosphate heptahydrate at 80 to 100, glucose at 1,000 to 2,000, EDTA-4 Na at 200 to 500, stem cells mild digestive enzyme at 1 to 120 and the balance of water. The mild digestive enzyme cell digestive juice for the stem cells, disclosed by the invention, has better digestion capacity on the cells than that of a traditional pancreatic enzyme, and the cells can be completely digested within 3 minutes. The mild digestiveenzyme cell digestive juice has less damage to the cells in a cell digestion process; after digestion is completed, the cells are inoculated, so that the cell viability is high and the growth state is good. The mild digestive enzyme cell digestive juice for the stem cells does not need to be stored at -20 DEG C, and can maintain the use effect at 2 to 8 DEG C within 12 months.

Description

Technical field [0001] The invention relates to a mild enzymatic digestion solution for stem cells, which is dedicated to the digestion of stem cells, and belongs to the related fields of biopharmaceuticals, biotherapy and the like. Background technique [0002] Trypsin is an animal-derived proteolytic enzyme, which is activated by trypsinogen secreted by the pancreas of animals. It can specifically hydrolyze the peptide bonds at the carboxyl end of arginine and lysine. It is widely used in the field of biotechnology and medical field. [0003] At present, there are two main ways to obtain trypsin. The traditional way is to obtain trypsin from animals. Trypsin is extracted from the pancreas by slaughtering cattle, sheep, pigs, etc. In addition, the use of microbial fermentation to obtain trypsin is gradually gaining popularity due to its high efficiency and specificity. [0004] With the continuous development of biopharmaceuticals, trypsin has become the main raw and auxiliary mat...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662
Inventor 李海峰张昕路明华闫攀登温郭秀王亚民
Owner YOCON BIOLOGY TECH CO
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