Cell preserving fluid for in-vitro analysis and detection and preparation method thereof

A technology for in vitro analysis and preservation solution, which is applied in the field of in vitro analysis and detection of cell preservation solution and its preparation, which can solve the problems of poor cell integrity, cell autolysis and corruption, and achieve enhanced protection, reduced autolysis and corruption, and good bactericidal effect Effect

Active Publication Date: 2022-01-25
深圳市华晨阳科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of cell preservation in the cell preservation solution on the market, the cells will still undergo some autolysis and corruption, which makes the integrity of cell preservation poor.

Method used

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  • Cell preserving fluid for in-vitro analysis and detection and preparation method thereof
  • Cell preserving fluid for in-vitro analysis and detection and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A cell preservation solution for in vitro analysis and detection, comprising components in the following concentration ranges: methanol 60v / v%, sodium chloride 0.54g / 100mL, potassium chloride 0.06g / 100mL, disodium edetate (hereinafter referred to as EDTA-2Na) 0.2g / 100mL, chitosan quaternary ammonium salt (substitution degree 90%) 0.3g / 100mL, trehalose 0.1g / 100mL; solvent is ultrapure water, pH 7.0.

[0036] Prepare 100 mL of the above cell preservation solution, including the following steps:

[0037] ① Take 20mL of ultrapure water, add 0.54g of sodium chloride, 0.06g of potassium chloride, 0.1g of disodium edetate, 0.3g of chitosan quaternary ammonium salt (substitution degree 90%) and 0.1g of trehalose g, stirred until completely dissolved to obtain the initial mixed solution;

[0038] ②. Add 63.2mL of 95% ethanol (approximately 60mL of ethanol content) to the initial mixture obtained in step ①, mix well and use 0.2M phosphate buffer (0.2mol / L NaH 2 PO 4 and 0.2mol...

Embodiment 2-6

[0041] Embodiments 2-6 are based on the method in Embodiment 1, and the component concentrations are adjusted, and the specific adjustments are shown in Table 1 below. Wherein, in Example 5, the component concentration of trehalose was adjusted to 0.05 g / mL, so that the weight concentration ratio of chitosan quaternary ammonium and salt trehalose was 6:1.

Embodiment 7-8

[0043] In Examples 7-8, on the basis of the method in Example 1, the types of cell fixatives were adjusted, and the details of the adjustments are shown in Table 1 below.

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Abstract

The invention relates to the technical field of biological preserving fluid, in particular to cell preserving fluid for in-vitro analysis and detection and a preparation method thereof. The cell preserving fluid is prepared from the following components in a concentration range: 60 to 75 v/v percent of a cell fixing agent, 0.6 to 0.9 g/100 mL of an osmotic pressure regulator, 0.2 to 0.5 g/100 mL of an anticoagulant, 0.3 to 0.6 g/100 mL of chitosan quaternary ammonium salt and 0.05 to 0.2 g/100 mL of trehalose; and the solvent is ultra-pure water, and the pH value is 7.0-7.6. A small amount of trehalose and chitosan quaternary ammonium salt are combined, so that the protective effect can be effectively enhanced, autolysis decay of cells is effectively reduced, and the integrity of cell preservation is improved. When the cell preserving fluid is prepared, ultra-pure water is used for dissolving components except the cell curing agent, the dissolving effect of the components can be effectively improved, the uniform and stable cell preserving fluid is obtained, and batch production of the cell preserving fluid is facilitated.

Description

technical field [0001] The present application relates to the technical field of biological preservation solution, in particular to a cell preservation solution for in vitro analysis and detection and a preparation method thereof. Background technique [0002] For cells obtained from non-blood sources such as the oral cavity, nasopharynx, and anus, how to maintain cell activity is a great challenge. When storing cells in a short period of time and applying them clinically, physiological saline is often used to maintain cell activity. However, the activity of cells in normal saline decreases rapidly, and the activity loss rate of cells can reach 30% or even higher within 10 hours. However, many observation tests on cells may take more than 10 hours or 24 hours from sampling to application. Even up to several days, saline can not meet the demand. [0003] In order to solve this problem, various cell preservation solutions have emerged as the times require. Cell preservation ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/021
Inventor 巩赞华巩赞博巩赞斌
Owner 深圳市华晨阳科技有限公司
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