Animal lung bud organoid culture medium and culture method

Abstract

The invention provides an animal lung bud organoid culture medium and culture method. The culture medium comprises a basic culture medium SAGM, sterile water and functional components, wherein the final concentration composition of the functional components in the organoid culture medium comprises: 0.5-2X of B27, 10-50 ng / ml of HGF, 2-20 ng / ml of BMP7, 20-200 ng / ml of EGF, 2-20 ng / ml of KGF, 100-400 ng / ml of FGF4, 1-20 [mu]M of SB216763, 10-100 nM of tretinoin, 20-200 ng / ml of Wnt-3A, 0.1-1 [mu]M of Prostaglandin E2, 0.5-2X of Insulin-Transferrin-Se, 5-20 mM of HEPES, 100 U / ml of penicillin, and 0.1 mg / ml of streptomycin. The culture medium of the invention is beneficial to the growth and function maintenance of lung bud organoids, and can culture lung tissues derived from mice and rats.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a culture medium and its application, and more specifically to a culture medium for animal lung bud organoids and a culture method. Background technique [0002] The lung is the respiratory organ of the living body, which realizes the gas exchange between the body and the external environment to maintain the life activities of the living body. The lung bud is the primitive structure of the embryonic lung. In the embryonic period, the terminal laryngotracheal diverticulum expands and develops into the budding tissue of the left and right lungs. The lung buds are formed by the gastrula during embryonic development. They are cord-like tissues in the early stage. Under the regulation of multiple factors and multiple pathways, the early lung buds gradually evolve into tubes, branching continuously, and forming pulmonary bronchial trees and alveoli. [0003] Lung bud organoids ha...

AI Technical Summary

Problems solved by technology

[0005] In the prior art, the culture of lung tissue cells is mainly cultured in a two-dimensional (2D) environment of monolayer cells. The interaction between cells makes the cultured lung tissue cells different from the living lung tissue cells, which is not conducive to the research

In 3D culture, in the absence of necessary and suitable medium, the culture and differentiation of lung tissue cells are also unfavorable, and it is difficult to fully simulate the physiological characteristics of the lung tissue structure in vivo

Although a variety of different culture conditions can be used to culture lung bud organoids derived from pluripotent stem cells or embryonic stem cells in vitro, there are no studies and reports on the culture methods of lung bud organoids derived from adult stem cells, especially specific experiments. There are not many reports on the process, operation steps, culture conditions and medium formulations

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