The invention provides a mouse
middle ear epithelium 3D
organoid and a construction method thereof, and is characterized in that the method comprises the following steps: a, taking a bubble structure of a mouse
middle ear tissue, cleaning, digesting, filtering, centrifuging filtrate, collecting precipitate, and separating primary cells of the mouse
middle ear; and b, inoculating the digested single cells and
cell agglomerates into a 24-pore plate, wrapping with
matrigel, and carrying out 3D culture, wherein a medium is a middle ear
epithelium amplification medium. And culturing for about 3-5 days to obtain the mouse 3D middle ear proliferation type
organoid. And performing differentiation culture by using a middle ear
epithelium differentiation medium to obtain the mouse middle ear epithelium organ. According to the method, the 3D mouse middle ear epithelium organ is successfully constructed, the physiological and
pathological processes of the middle ear are conveniently analyzed and researched, the interaction between the middle ear microenvironment and cells can be more accurately simulated, and the application range is wide as a model.