Nasal mucosa organoid culture medium and culture method
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A culture medium and nasal mucosa technology, which is applied in the field of organoid culture, can solve the problems of no attempt and report on the formulation of the experimental process, the culture conditioned medium of the operation steps, and no research and report on the culture method of nasal mucosa tissue organoids, and achieves success. High rate, high repeatability, technically stable effect
Active Publication Date: 2020-05-08
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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Problems solved by technology
Although a variety of human-derived tissues can successfully culture primary cells or organoids in vitro using different methods and under different culture conditions, there is currently no research on the culture method
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Embodiment 1
[0048] A nasal mucosa organoid culture medium, its composition and content are as follows: B27 without Vit-A, 1X; N-acetylcysteine, 0.2mM; EGF, 10ng / ml; Noggin, 20ng / ml; R-spondin 1, 50ng / ml; Wnt3a, 50ng / ml; A83-01, 100nM; Nicotinamide, 2mM; Y-27632, 2μM; Glutamax, 1X; Gastrin, 1nM; SB202190, 2μM; / ml; IL-6, 10 ng / ml; HEPES, 0.2 mM.
Embodiment 2
[0050] A nasal mucosal organoid culture medium, its composition and content are as follows: B27 without Vit-A, 5X; N-acetylcysteine, 5mM; EGF, 250ng / ml; Noggin, 500ng / ml; R-spondin 1, 1000ng / ml; Wnt3a, 1000ng / ml; A83-01, 2500nM; Nicotinamide, 50mM; Y-27632, 50μM; Glutamax, 5X; Gastrin, 25nM; SB202190, 50μM; ml; IL-6, 250 ng / ml; HEPES, 5 mM.
Embodiment 3
[0052] A nasal mucosal organoid culture medium, its composition and content are as follows: B27 without Vit-A, 1X; N-acetylcysteine, 1mM; EGF, 50ng / ml; Noggin, 100ng / ml; R-spondin 1, 250ng / ml; Wnt3a, 250ng / ml; A83-01, 500nM; Nicotinamide, 10mM; Y-27632, 10μM; Glutamax, 1X; Gastrin, 5nM; SB202190, 10μM; ml; IL-6, 50 ng / ml; HEPES, 1 mM.
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Abstract
The invention discloses a culture medium and culture method of a nasal mucosa tissue. According to the culture medium and culture method of the nasal mucosa tissue, epithelial-mesenchymal/matrix components are integrated in an organoid culture system based on a gas-liquid interface method for the first time, and by inducing proliferation and differentiation of adult stem cells in a fresh nasal mucosa epithelial tissue, an organoid which is composed of multiple kinds of cells, including ciliated cells, goblet cells, club cells and basal cells, and is close to an internal mucosa in structure andfunction can be obtained, and becomes an in-vitro nasal mucosa model with proliferation and multi-lineage differentiation capability.
Description
technical field [0001] The present invention relates to the field of organoid culture, further relates to the field of nasal mucosa organoid culture, in particular to a nasal mucosa organoid culture medium and a culture method. Background technique [0002] Organoids are organ-specific collections of cells derived from stem cells or precursor cells. Organoids cultured in vitro are highly similar to corresponding organs in terms of cellular components and tissue architecture, and have corresponding functional characteristics. Organoid culture represents a major revolution in conventional cell biology techniques. Unlike conventional cell culture, which cultivates a single cell group in a two-dimensional environment, organoid culture is to cultivate a variety of cell groups contained in specific tissues and organs in a three-dimensional environment, and its culture system is more similar to the microenvironment in vivo. Therefore, it has shown great application prospects in b...
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