Method for efficiently inducing advanced branched pulmonary organs in vitro, experimental model and compound combination

Abstract

The invention provides a method for efficiently inducing advanced branched pulmonary organs in vitro, an experimental model and a compound combination, and relates to the technical field of biology. The method is to digest airway epithelial tissues in vitro, then carrying out multiplication culture on the airway epithelial cells by utilizing an amplification culture medium containing epidermal growth factors and insulin, digesting the airway epithelial cells into single cells by utilizing pancreatin, and then adding the single cells into a differential culture medium for culture to obtain a mouse lung organ which has a branched advanced structure and contains differentiated ciliated cells. The mouse lung organ can be used as an experimental model for precision medical treatment, organ transplantation, drug screening and mechanism research on drug action. According to the method disclosed by the invention, the airway epithelial cells can be cultured into the lung organs which have branched advanced structures and differentiated ciliated cells that are more similar to the lung morphology and cell structure in vivo, and the yield of the cultured lung organs is more than three times that of the lung organs cultured by a conventional method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for efficiently inducing high-level branched lung organoids in vitro, an experimental model and a combination of medium used. Background technique [0002] Lung organoids are small tissues similar to lungs in structure and function established by in vitro culture technology. They have the characteristics of tissue self-renewal and long-term culture. To a certain extent, they simulate the physiological activities and pathological changes of lungs in vivo. Ideal in vitro carrier for organ transplantation, drug screening, and drug mechanism research. [0003] Lung organoids can be generated by inducing human pluripotent stem cells or mouse adult stem cells in 3D media supplemented with lung-associated growth factors. In 2015, scientists for the first time obtained the first lung organoids from human stem cells (see reference 1). In 2017, Professor Hans-Willem Snoeck and colle...

AI Technical Summary

Problems solved by technology

Such culture methods can form pseudostratified structures resembling lung airway epithelial cells, but without the luminal morphology and structure specific to in vivo airway tissue

②Cultivate the isolated basal cells under the 3D condition of Matril gel without growth factors, and induce differentiation to produce spherical lung organoids (see references 6-7). Compared with method ①, this culture method can form and An organization more similar to the tubular tissue of the lung airways, but without the higher branching structure of the lung airway tissue in vivo

In addition, due to the lack of airway basal cells in mice and the harsh culture environment for organoid formation, the existing culture methods all have the problem of low organoid yield.

[0004] In summary, although lung organoids formed in vitro can be widely used in basic research and drug testing, there are still The low yield and lack of complex branched lumen structure in the lung severely limit the application of lung organoids

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