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Preserving fluid and preserving method for keeping cell activity at low temperature

A low-temperature condition and cell activity technology, applied in the biological field, to achieve simple operation steps, ensure accuracy, and solve the effects of preservation and transportation

Inactive Publication Date: 2020-08-11
上海伯豪生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] One of the technical problems to be solved in the present invention is to solve the storage and transportation of cell samples after collection, maintain the activity of cells under low temperature conditions, and do not need steps such as cryopreservation and recovery, and do not add components that cause changes in cell gene expression

Method used

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  • Preserving fluid and preserving method for keeping cell activity at low temperature
  • Preserving fluid and preserving method for keeping cell activity at low temperature
  • Preserving fluid and preserving method for keeping cell activity at low temperature

Examples

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Embodiment 1

[0028] Embodiment 1: the preparation of preservation solution A

[0029] In a specific embodiment, the composition of the preservation solution includes potassium hydroxide (5-6 g / L), lactobionic acid (30-40 g / L), sodium dihydrogen phosphate (3-5 g / L), Magnesium sulfate water (0.5-1 g / L), raffinose (15-20 g / L), hydroxyethyl starch (50-60 g / L), adenosine (1-2 g / L), allopurinol (0.2-0.5 g / L), glutathione (0.5-2 g / L), dissolved in sterilized deionized water, using sodium hydroxide and dilute hydrochloric acid to adjust the pH of the solution to 7.2-7.6.

Embodiment 2

[0030] Embodiment 2: the preparation of preservation solution B

[0031] In a specific embodiment, the composition of the preservation solution includes potassium hydroxide (1-1.5 g / L), potassium chloride (0.8-1.2 g / L), 4-hydroxyethylpiperazineethanesulfonic acid (5- 6 g / L), lactobionic acid (30-40 g / L), dextran (50-60 g / L), magnesium chloride (0.6-1.2 g / L), sodium dihydrogen phosphate (1-2 g / L ), sodium bicarbonate (0.4-0.6 g / L), glucose (0.5-1 g / L), sucrose (5-7 g / L), mannose (4-5 g / L), adenosine (1- 2 g / L), glutathione (0.5-2 g / L), vitamin E (0.2-0.5 g / L), dissolved in sterilized deionized water, using sodium hydroxide and dilute hydrochloric acid to adjust the pH of the solution to 7.2-7.6.

Embodiment 3

[0032] Embodiment 3: the preparation of preservation solution C

[0033] In a specific embodiment, the composition of the preservation solution includes potassium hydroxide (1-1.5 g / L), potassium chloride (0.8-1.2 g / L), 4-hydroxyethylpiperazineethanesulfonic acid (5- 6 g / L), lactobionic acid (30-40 g / L), dextran (50-60 g / L), magnesium chloride (0.6-1.2 g / L), potassium dihydrogen phosphate (1-2 g / L ), sodium bicarbonate (0.4-0.6 g / L), glucose (0.5-1 g / L), sucrose (5-7 g / L), mannose (4-5 g / L), adenosine (1- 2g / L), allopurinol (0.2-0.5 g / L), glutathione (0.5-2 g / L), dissolved in sterilized deionized water, using sodium hydroxide and dilute hydrochloric acid to adjust the pH of the solution to 7.2-7.6.

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Abstract

The invention provides a preserving fluid and a preserving method for keeping cell activity under a low-temperature condition. A formula for the preserving fluid is optimized to enable the componentsof the preserving fluid to maintain the activity of cells at a low temperature. The pH value of the preserving fluid is 7.2-7.6; and a preservation temperature is 0-6 DEG C. The components do not contain components which are toxic to cells and cause gene expression change, so the preserving fluid is safe and environment-friendly. Meanwhile, cryopreservation, recovery and other processes are not needed in the preserving method, cells are kept in an active state in the whole process, and the method is simple and easy to use, and facilitates cell storage and transportation. According to the invention, low-temperature storage of various types of human and animal cells can be effectively realized, the activity of the cells can be maintained under a low-temperature condition, and it is verifiedthat the preserving fluid can be used for low-temperature storage and transportation of human blood cells, cultured cells and tissue sample digested and dissociated cells. The preserved cells can be used for gene detection related research or other cell preservation and other related application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preservation solution for maintaining the activity of human or animal cells under low temperature conditions; in addition, the invention also relates to a preservation method using the preservation solution. Background technique [0002] With the gradual deepening of relevant research on genetic testing technology and the popularization of clinical applications, how to safely and effectively preserve biological samples has become the key. Among them, the most widely studied cell samples, such as clinical blood cells, tissue digested and dissociated cells, and cultured cells, have time and space distances from collection to laboratory testing. How to ensure that cell samples are maintained during storage and transportation The stability of cell activity and gene expression will determine whether the experimental results are accurate and effective. In particular, the single-cell seq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 刘崇懿李婷婷孟宪欣肖华胜
Owner 上海伯豪生物技术有限公司
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