Culture method of mouse lung organs and special culture solution thereof

Abstract

The invention discloses a culture method of mouse lung organs and a special culture solution thereof. The method comprises the following steps: acquiring fresh mouse lung tissue cells, digesting into single cells by collagenase, culturing mouse lung tissue organoid under an in-vitro three-dimensional culture condition, and performing immunofluorescent staining on the lung organoid. The culture solution is prepared from ADF < + + + + >, an RPSO1 conditional culture medium, a Noggin conditional culture medium, a B27 additive, N-acetyl-L-cysteine, nicotinamide, Y-27632, A83-01, SB202190, a recombinant human fibroblast growth factor-7 and a recombinant human fibroblast growth factor-10. Experiments show that the lung organ established by the method disclosed by the invention can effectively maintain tissue cell specificity, stem cell characteristics and biological functions, and meets the requirements of scientific research.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for culturing mouse lung organoids and a special culture solution thereof. Background technique [0002] Organoids are microscopic tissue and organ analogs formed by three-dimensional culture in vitro by utilizing the self-renewal and differentiation capabilities of stem cells, which to a large extent have the functions of corresponding organs in vivo. The three-dimensional culture of organoids requires the use of bioengineering methods to guide cell division and differentiation. Cytokines and extracellular matrix constitute the microenvironment of stem cell culture, which is the material basis for organoid renewal and differentiation. By artificially regulating the components of the culture system, the cells are autonomously differentiated into specific structures, and the organoid self-assembly process is completed. At present, organoids of esophagus, stomach...

AI Technical Summary

Problems solved by technology

Two-dimensionally cultured lung tissue cells cannot be passaged for a long time, and genotype and phenotype changes will gradually occur during the passage process; limited by two-dimensional culture conditions, it is difficult to fully express the characteristics of lung tissue, which is different from living lung tissue cells. discrepancy, which is not conducive to the conduct of research

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