Universal culture mediums and culture method for upper respiratory tract mucosa organs

An upper respiratory tract and organoid technology, which is applied in the medium and culture field of general-purpose upper respiratory tract mucosal organoids, can solve the problem of experimental procedures, operation steps, and the formulation of culture conditioned medium without too many reports, and upper respiratory tract mucosal tissue organoids. There are few problems in the research and reports of culture methods, which are beneficial to the maintenance of growth and function, have a wide range of applications, and improve the growth rate.

Active Publication Date: 2020-07-31
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although a variety of other human tissues (such as liver, intestine, etc.) can be successfully cultured into organoids in vitro using different methods and under different culture conditions, the current research and reports on the culture methods of upper respiratory mucosa tissue organoids Fewer, especially the specific test process, operation steps, culture conditions, that is, there are not many reports on the medium formula

Method used

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  • Universal culture mediums and culture method for upper respiratory tract mucosa organs
  • Universal culture mediums and culture method for upper respiratory tract mucosa organs
  • Universal culture mediums and culture method for upper respiratory tract mucosa organs

Examples

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Embodiment 1

[0035] This embodiment provides a general-purpose liquid medium for upper respiratory tract mucosal organoids, including basic cytokines, specific cytokines, inhibitors, penicillin, streptomycin, sterile water and BEGM culture medium, wherein the basic cells Factors include: 50ng / ml of EGF, 200ng / ml of Noggin, 250ng / ml of R-spondin 1, 300ng / ml of Wnt3a; the specific cytokines include: 20ng / ml of Wnt7a, 100ng / ml of IL- 6. The inhibitors include: 500nM EW-7197-HCl, 10μM SB 203580-HCl, 20μM Y-27632dihydrochloride; the penicillin concentration is 100U / ml; the streptomycin concentration is 0.1mg / ml; The concentration of each of the above components is based on the concentration in the mixture of BEGM culture solution and sterile water.

Embodiment 2

[0037]This embodiment provides a general-purpose liquid medium for upper respiratory tract mucosal organoids, including basic cytokines, specific cytokines, inhibitors, penicillin, streptomycin, sterile water and BEGM culture medium, wherein the basic cells Factors include: 20ng / ml of EGF, 400ng / ml of Noggin, 200ng / ml of R-spondin 1, 100ng / ml of Wnt3a; the specific cytokines include: 10ng / ml of Wnt7a, 50ng / ml of IL- 6. The inhibitors include: 200nM EW-7197-HCl, 20μM SB 203580-HCl, 40μM Y-27632dihydrochloride; the penicillin concentration is 100U / ml; the streptomycin concentration is 0.1mg / ml; The concentration of each of the above components is based on the concentration in the mixture of BEGM culture solution and sterile water.

Embodiment 3

[0039] This example provides a general-purpose gel medium for upper respiratory tract mucosal organoids, including basic cytokines, specific cytokines, inhibitors, penicillin, streptomycin, Matrigel matrigel, sterile water and BEGM culture medium, Wherein the basic cytokines include: 50ng / ml EGF, 200ng / ml Noggin, 250ng / ml R-spondin 1, 300ng / ml Wnt3a; the specific cytokines include: 20ng / ml Wnt7a, 100ng IL-6 per ml; the inhibitors include: 500nM EW-7197-HCl, 10μM SB 203580-HCl, 20μM Y-27632dihydrochloride; the penicillin concentration is 100U / ml; the streptomycin concentration is 0.1 mg / ml; the concentration of Matrigel is 5 mg / ml; the concentration of each of the above components is based on the concentration in the mixture of BEGM culture medium and sterile water.

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Abstract

The invention provides universal culture mediums and culture method for upper respiratory tract mucosa organs. A liquid culture medium comprises a basic cell factor, a specific cell factor, an inhibitor, penicillin, streptomycin, sterile water and a BEGM culture solution. A gel culture medium is further added with Matrigel with a concentration of 3-6 mg/ml on the basis of the liquid culture medium. The culture mediums are used for culturing upper respiratory tract mucosa tissue organs, are wide in application range, can be used for culturing tissues from various sample sources such as noses, mouths and throats, remarkably improve culture effect, mucosa tissue cell differentiation and cell cilium growth speed, and are beneficial to growth and function maintenance of upper respiratory tractmucosa tissue cells.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a culture medium and a culture method for general-purpose upper respiratory tract mucosal organoids. Background technique [0002] The upper respiratory tract mainly includes the nose, pharynx, and larynx. Secretion and cilia are distributed on the inner surface of the upper respiratory tract (except nostrils, posterior pharyngeal wall and vocal cord mucosa), which can warm (or cool), moisten and purify the inhaled air, and have a protective effect on respiratory organs and the human body. The upper respiratory tract plays a very important role in filtering and cleaning the human body. Through the filtering and cleaning function of the respiratory tract, foreign objects such as particles and pathogens that enter the respiratory tract with the air are blocked and removed, and the gas entering the alveoli is almost clean and sterile. These functions are mainly It i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0697C12N2501/11C12N2501/2306C12N2501/415C12N2501/999
Inventor 李刚汪珂陈泽新
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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