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Cell fixing agent and cell fixing method

A cell fixation and cell technology, applied in the preservation, application, animal husbandry and other directions of human or animal body, can solve problems such as harm and toxicity, and achieve the effects of high antigenic activity, strong mechanical properties, and convenient recovery.

Pending Publication Date: 2022-04-01
苏州良辰生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the patent CN110973117B reduces the amount of toxic substances used to a certain extent, it is safer and more environmentally friendly than traditional cell fixation reagents, but it still cannot avoid the use of alcohols and aldehydes, so it still has certain toxicity and hazards

Method used

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  • Cell fixing agent and cell fixing method
  • Cell fixing agent and cell fixing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Fixation of PG-4 cells (astrocytes)

[0058] (1) Fixative configuration: Weigh 0.30g of polyvinylamine, 0.20g of vitamin C and 0.02g of diatomaceous earth, dissolve them in 10mL of PBS buffer, and mix well.

[0059] (2) PG-4 cell culture: Take a 6-well cell culture plate and add 1.5×10 5 pcs / hole (5×10 4 cells / mL) of PG-4 cells, supplemented with culture medium (McCoy’s 5A medium containing 10% fetal bovine serum) to 3mL per well and placed at 37°C, containing 5% CO 2 Incubate overnight in the incubator. The next day, the cell culture plate was taken out, the culture medium was discarded, and each well was washed twice with serum-free McCoys'5A medium.

[0060] (3) Cell fixation: 0.5 mL of fixative solution was added to each well of the above-mentioned cell culture, and the cells were fixed at room temperature for 4 hours, and the fixative solution was discarded. Stain with Gimsa staining solution (Gimsa) for 5 minutes, discard the staining solution, wash...

Embodiment 2

[0061] Example 2: Fixation of Pseudomonas aeruginosa

[0062] (1) Thalline preparation: Pseudomonas aeruginosa was activated and cultivated to obtain bacterial liquid, and the bacterial liquid was centrifuged to obtain wet bacterial cells. Dissolve 0.5 g of wet cells in 10 mL of PBS pre-cooled at 2-8°C, and mix well to make a cell suspension.

[0063] (2) Add 0.30 g of polyethyleneimine, 0.20 g of vitamin C and 0.04 g of diatomaceous earth in sequence to the cell suspension, mix well, and leave to stand at room temperature for 2 hours for cross-linking.

[0064] (3) The solid and liquid were separated by vacuum filtration, and the fixed cells were washed three times with PBS buffer solution, and set aside.

[0065] (4) Dissolve the prepared immobilized cells in PBS buffer and shake with glass beads for 24 hours. Use the human β-lactamase (LACTβ) kit to detect the residual enzyme activity of β-lactamase after cell fixation, and take the same amount at the same time. The initi...

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Abstract

The invention provides a cell fixing agent and a cell fixing method. In order to solve the problems that volatile organic substances need to be used in an existing cell curing agent, harm is caused to a human body and the environment, and meanwhile, in order to improve the mechanical strength of cured cells and the activity of antigens in the cells, the cell fixing agent provided by the invention comprises a solvent, effective components and a fixing carrier, the effective components comprise polyethyleneimine and vitamin C, and the solvent is a PBS buffer solution and / or normal saline. According to the present invention, by adjusting the formula and the ratio of the cell fixing agent, the cell fixing effect is ensured, the volatile organic substance is not used, the harm to the user and the influence on the environment are reduced, and the cell subjected to the immobilization treatment has characteristics of strong mechanical property, convenient recovery, high antigen activity in the cell, and good stability. And high-strength antigen production is facilitated.

Description

technical field [0001] The invention relates to the technical field of cell fixation, in particular to a cell fixative and a cell fixation method. Background technique [0002] Immobilized cell technology is to fix free cells on a specific carrier by physical or chemical methods, so that they can be reused and maintain their vitality. Immobilized cells have a wide range of applications and have been used in various fields such as industry, medicine, pharmacy, chemical analysis, environmental protection, and energy development. [0003] The key to cell fixation is to choose a suitable cell fixative to keep the cell shape unchanged, while keeping the three-dimensional structure of the antigen unchanged, and avoiding the inactivation or diffusion of the antigen. Currently commonly used cell fixatives mainly include: aldehydes (commonly used formaldehyde, glutaraldehyde and paraformaldehyde), alcohols (commonly used ethanol) and ketones (such as acetone). These fixatives must ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 吴娟
Owner 苏州良辰生物医药科技有限公司
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