Method for rapidly separating and culturing human bronchial epithelial cells, and optimized medium

Abstract

The invention discloses a method for rapidly separating and culturing human bronchial epithelial cells, and an optimized medium. The optimized medium is prepared from a DMEM culture medium, an F-12 Nutrient mix culture medium, 0.5 mg/ml hydrocortisone, fetal bovine serum, 25mu g/ml epidermal growth factor, 5 mg/ml insulin, 11.7mu M cholera toxin, 10 mM ROCK1 inhibitor, and 10 mM BMP4 antagonist. The method for rapidly culturing the human bronchial epithelial cells by using the optimized medium not only greatly simplifies the steps of culturing epithelial cells which require 3T3 J2 cells as a feeder layer, and only needs to culture the primary human bronchial epithelial cells digested by pronase in the optimized medium, but also can be used for obtaining humane bronchial epithelial cells which are rapidly separated and can be indefinitely subcultured; the subcultured human bronchial epithelial cells are subjected to in vitro expansion and culture; when the human bronchial epithelial cells are subcultured for 20 generations or more, a sufficient amount of seed cells can be obtained so as to be used in a variety of follow-up in vitro and in vivo studies such as in vitro cell interaction model studies.

Description

Technical field: [0001] The invention belongs to the technical field of primary cell separation and culture, and in particular relates to a method for rapidly separating and culturing human airway epithelial cells and an optimized culture medium. Background technique: [0002] The human airway is mainly a pseudostratified epithelium composed of three types of epithelial cells, including basal cells, goblet cells, and ciliated cells. The basal cells are arranged in the basement membrane and are currently recognized as airway epithelial stem cells. Goblet cells produce Mucus traps inhaled particles and pathogens, while ciliated cells generate motility to remove them from the lungs. Impaired mucociliary clearance is an important challenge after tracheal transplantation because secretions are retained at the distal anastomotic site. The in vitro culture and proliferation of airway epithelial cells is the basis of airway epithelial function and regenerative medicine research, bu...

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Problems solved by technology

However, the operation is complicated and cumbersome, and the cell state is great

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