Method for establishing adult mouse cochlea body culture system and application thereof

Abstract

The invention relates to a method for establishing an adult mouse cochlea body culture system and application thereof. According to the invention, a co-culture structure of inner ear sensory epithelium and spiral ganglion neurons of an adult mouse, which are provided with a bony scaffolds, is placed into culture solution to carry out culture, and the culture solution is regularly replaced; and components of the culture solution comprises N2, B27 and penicillin. According to the invention, a first integrated culture system of the cochlea sensory epithelium and the spiral ganglion neurons of an adult mammal is established; completeness of an inner ear sensory epithelium structure and the survival of sustentacular cells and the spiral ganglion neurons can be maintained for a long term, a research model is provided for research on proliferation and transdifferentiation of the inner ear sustentacular cells, differentiation and maturation of hair cells, establishment of synaptic connection between newborn hair cells and the spiral ganglion neurons and the like for the adult mammals, the blank of models in the field is filled, and the research level reaches the international leading level.

Description

technical field [0001] The invention belongs to the technical field of in vitro cochlear culture, and in particular relates to a method for establishing an adult mouse cochlear body culture system and its application. Background technique [0002] According to the statistics of the World Health Organization in 2019, there are currently about 466 million hearing-impaired patients in the world, accounting for more than 5% of the total population. More than 30% of people have disabling hearing impairment, which is a major disease that seriously threatens human health. The current study believes that the irreversible damage and loss of hair cells, the key component of the inner ear receptors that play the role of mechanical-electrical conversion, is the most important cause of sensorineural deafness. At the same time, mammalian cochlear hair cells cannot spontaneously Regeneration is key to making deafness hard to cure. [0003] The organ of Corti in mammals is composed of sup...

AI Technical Summary

Problems solved by technology

The in vitro culture system of neonatal mouse cochlea sensory epithelium has been widely used in the study of hair cell regeneration, and a series of regulatory factors that can promote hair cell regeneration have been screened out. However, due to the lack of in vitro culture system for adult mouse cochlea, these regulatory factors The effectiveness of factors promoting hair cell regeneration in the adult stage cannot be verified efficiently, and the vast majority of patients faced by the present invention are adults or even the elderly, and the effective regeneration of hair cells in mammals after adulthood is important for sensorineural The treatment of deafness has greater value, so it is urgent to establish an in vitro culture system for adult mouse cochlea, and quickly and efficiently screen adult hair cell regeneration strategies

[0005] In the traditional cochlear basement membrane culture of neonatal mice, the basement membrane is completely separated from the bony structure and attached to the bottom of the culture dish, which can maintain tissue activity for a long time in a suitable culture system, but the cochlea is severely ossified after adulthood Along with the reduction of the toughness of the sensory epithelium, structural damage is inevitable during the separation of the basement membrane. In addition, due to the decrease in the number and proliferation of fibroblasts in the basement membrane, they cannot stretch and adhere well in culture. The bottom of the dish is prone to folding and curling, further aggravating the damage of basement membrane cells, so most of the hair cells and supporting cells will be lost during the long-term culture process, making it impossible to carry out relevant research work

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