Reagent for eliminating mycoplasma contamination in cell culture and use method thereof

A technology for removing cells and mycoplasma, which is applied in tissue culture, biochemical equipment and methods, and microbial measurement/inspection, and can solve the problems of reagents and methods that have not been studied for mycoplasma contamination.

Active Publication Date: 2012-04-11
胡晓鹏
View PDF4 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the classification of mycoplasma is different from that of bacteria, the antibiotics generally used for bacteria cannot kill and inhibit mycoplasma very well. Due to the variety of mycoplasma, it is necessa

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reagent for eliminating mycoplasma contamination in cell culture and use method thereof
  • Reagent for eliminating mycoplasma contamination in cell culture and use method thereof
  • Reagent for eliminating mycoplasma contamination in cell culture and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047]The reagent for removing mycoplasma contamination in cell culture is formed by mixing equal volumes of liquid A and liquid B, wherein:

[0048] Liquid A is formed by mixing tiamulin and 0.01MPBS, and the mass-volume concentration of tiamulin in the mixture is 1.5%;

[0049] Solution B is formed by mixing enrofloxacin and 0.01MPBS, and the mass-volume concentration of enrofloxacin in the mixed solution is 1%;

[0050] Mix equal volumes of liquid A and liquid B, and the mass ratio of tiamulin to enrofloxacin in the mixed liquid is 3:2.

[0051] A method for removing mycoplasma contamination with the above reagents, selects the cultivation of A549 cells (human lung adenocarcinoma cells) as the experimental object, and its concrete steps are as follows:

[0052] a. Mycoplasma detection

[0053] (1) The cell culture medium of A549 cells was prepared by adding 10% FBS (domestic Sijiqing fetal bovine, mycoplasma-free grade) into DMEM high-glucose medium (manufactured by Ameri...

Embodiment 2

[0069] The reagent for removing mycoplasma contamination in cell culture is formed by mixing equal volumes of liquid A and liquid B, wherein:

[0070] Liquid A is formed by mixing vornemulin and 0.01MPBS, and the mass-volume concentration of vornemulin in the mixed solution is 1.5%;

[0071] Liquid B is formed by mixing marbofloxacin and 0.01MPBS, and the mass-volume percentage concentration of marbofloxacin in the mixed solution is 1%;

[0072] Liquid A and liquid B are mixed in equal volumes, and the mass ratio of warnemulin to marbofloxacin in the mixed liquid is 3:2.

[0073] A kind of method that removes mycoplasma pollution with above reagent, selects the cultivation of SH-SY-5Y cell (human neuroblastoma) as experiment, and its concrete steps are as follows:

[0074] a. Mycoplasma detection

[0075] (1) For the cell culture medium of SH-SY-5Y cells, DMEM high-glucose medium (manufactured by GIBCO) and 10% FBS (domestic Sijiqing Tainiu, mycoplasma-free grade) were added...

Embodiment 3

[0092]The reagent for removing mycoplasma contamination in cell culture is formed by mixing equal volumes of A liquid, B liquid, and C liquid, wherein:

[0093] Liquid A is formed by mixing vornemulin and 0.01MPBS, and the mass-volume concentration of vornemulin in the mixed solution is 1.5%;

[0094] Liquid B is formed by mixing enrofloxacin and 0.01MPBS, and the mass-volume percentage concentration of enrofloxacin in the mixed liquid is 1%;

[0095] Liquid C is formed by mixing minocycline and 0.01MPBS, and the mass-volume percentage concentration of minocycline in the mixed solution is 0.5%;

[0096] Liquid A, liquid B, and liquid C are mixed in equal volumes, and the mass ratio of warnemulin, enrofloxacin, and minocycline in the mixed liquid is 3:2:1.

[0097] A method for removing mycoplasma contamination with the above reagents, selects the cultivation of HELA cells as an experiment, and its concrete steps are as follows:

[0098] a. Mycoplasma detection

[0099] (1) ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a reagent for eliminating mycoplasma contamination in cell culture and a use method thereof. The reagent is prepared by mixing liquid A and liquid B isometrically or mixing liquid A, liquid B and liquid C isometrically, wherein the liquid A is prepared by mixing semisynthetic pleuromutilin derivatives and 0.01MPBS, and the mass-volume percentage concentration of the solute after mixing is 0.5-1.5%; the liquid B is prepared by mixing quinolone derivatives and 0.01MPBS, and the mass-volume percentage concentration of the solute after mixing is 0.5-1.5%; and the liquid C is prepared by mixing tetracycline derivatives and 0.01MPBS, and the mass-volume percentage concentration of the solute after mixing is 0.25-0.75%. The reagent and method provided by the invention havethe following beneficial effects: if contamination is not severe, the effect can be displayed within a week; and after the treatment cycle is finished, black spots do not exist among the treated contaminated cells, the growth cycles of the cells return to normal, the cell morphology is normal, and aggregated growth and netting phenomena are avoided.

Description

technical field [0001] The invention relates to a reagent for removing mycoplasma contamination in cell culture and a method for using the same. Background technique [0002] Mycoplasma is the most common pollutant in cell culture, which will interfere with the experimental results. However, since the binary division time of mycoplasma is 1-6 hours, it is difficult to detect the initial contamination of cultured cells by mycoplasma. Common contaminating mycoplasmas in cell cultures were derived from M. orale, M. arginini, M. hyorhinis and A. laidlawii. After the cultured cells are contaminated with mycoplasma, they will have different pathogenicity according to the antigenicity and biochemical metabolism of different mycoplasmas, but the general performance is the same. At first, the experimenter noticed that the cultured cells grew slowly, and then the culture medium was very slow. It will soon turn yellow, and the cell growth cycle will be prolonged, and then a large numb...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/00C12Q1/68C12Q1/04
Inventor 胡晓鹏
Owner 胡晓鹏
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap