The present invention provides a method for prolonging the
in vitro culture time of cochlear hair cells, belonging to the field of
cell culture, and the method steps are as follows: (1) dissecting the
temporal bone: peeling off the mouse forebrain,
cerebellum and
brainstem, and separating and obtaining the
cochlea; (2) extracting the
cochlea: separating the spiral
ligament on the Corti organ from the rest of the
cochlea, peeling off the Reissner's membrane, and removing the
tectorial membrane above the Corti organ; (3) culturing the cochlea
in vitro: placing the cochlear hair cells upward on a
cell culture plate containing a
cell slide and a culture medium, wherein the cochlear
hair cell culture medium is a DMEM high-glucose culture medium containing 0.6-0.7% glucose (including glucose in the basal culture medium), 1% N2 supplement and 100U / mL
penicillin G
sodium, and culturing the hair cells in a
cell culture incubator, and replacing the culture medium every 72 hours. The
in vitro culture method of cochlear hair cells provided by the present invention is the first reported method at home and abroad, and can culture hair cells in vitro for 30 days, which can meet the needs of exploring the mechanism of
hair cell development and differentiation and collecting data on in vitro
gene therapy for hearing impairment.