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Abstract: Auditory function in vertebrates depends on the transduction of sound vibrations into electrical signals by inner ear hair cells. In general, hearing loss resulting from hair cell damage is irreversible because the human ear has been considered to be incapable of regenerating or repairing these sensory elements following severe injury. Therefore, regeneration and protection of inner ear hair cells have become an exciting, rapidly evolving field of research during the last decade. However, mammalian auditory hair cells are few in number, experimentally inaccessible, and barely proliferate postnatally in vitro. Various in vitro primary culture systems of inner ear hair cells have been established by different groups, although many challenges remain unresolved. Here, we briefly explain the structure of the inner ear, summarize the published methods of in vitro hair cell cultures, and propose a feasible protocol for culturing these cells, which gave satisfactory results in our study. A better understanding of in vitro hair cell cultures will substantially facilitate research involving auditory functions, drug development, and the isolation of critical molecules involved in hair cell biology.

哺乳动物内耳毛细胞的体外培养

[1]Abdouh A, Despres G, Romand R, 1993. Hair cell overproduction in the developing mammalian cochlea in culture. Neuroreport, 5(1):33-36.

[2]Anniko M, van de Water TR, 1978. Organ culture of the postnatal mouse crista ampullaris part I. Arch Oto-Rhino-Laryngol, 220(1-2):129-132.

[3]Betlejewski S, 2008. Science and life—the history of marquis Alfonso Corti. Otolaryngol Pol, 62(3):344-347.

[4]Bird JE, Daudet N, Warchol ME, et al., 2010. Supporting cells eliminate dying sensory hair cells to maintain epithelial integrity in the avian inner ear. J Neurosci, 30(37):12545-12556.

[5]Brandon CS, Voelkel-Johnson C, May LA, et al., 2012. Dissection of adult mouse utricle and adenovirus-mediated supporting-cell infection. J Vis Exp, (61):e3734.

[6]Bucks SA, Cox BC, Vlosich BA, et al., 2017. Supporting cells remove and replace sensory receptor hair cells in a balance organ of adult mice. eLife, 6:e18128.

[7]Chang DT, Chai R, DiMarco R, et al., 2015. Protein-engineered hydrogel encapsulation for 3-D culture of murine cochlea. Otol Neurotol, 36(3):531-538.

[8]Corns LF, Johnson SL, Kros CJ, et al., 2014. Calcium entry into stereocilia drives adaptation of the mechanoelectrical transducer current of mammalian cochlear hair cells. Proc Natl Acad Sci USA, 111(41):14918-14923.

[9]Ding DL, Stracher A, Salvi RJ, 2002. Leupeptin protects cochlear and vestibular hair cells from gentamicin ototoxicity. Hear Res, 164(1-2):115-126.

[10]Ding DL, He JC, Allman BL, et al., 2011. Cisplatin ototoxicity in rat cochlear organotypic cultures. Hear Res, 282(1-2):196-203.

[11]Ding DL, Qi WD, Yu DZ, et al., 2013. Addition of exogenous NAD⁺ prevents mefloquine-induced neuroaxonal and hair cell degeneration through reduction of caspase-3-mediated apoptosis in cochlear organotypic cultures. PLoS ONE, 8(11):e79817.

[12]Dror AA, Avraham KB, 2009. Hearing loss: mechanisms revealed by genetics and cell biology. Annu Rev Genet, 43:411-437.

[13]Eatock RA, 2000. Adaptation in hair cells. Annu Rev Neurosci, 23:285-314.

[14]Friedman TB, Griffith AJ, 2003. Human nonsyndromic sensorineural deafness. Annu Rev Genomics Hum Genet, 4:341-402.

[15]Furness DN, Richardson GP, Russell IJ, 1989. Stereociliary bundle morphology in organotypic cultures of the mouse cochlea. Hear Res, 38(1-2):95-109.

[16]Gaboyard S, Chabbert C, Travo C, et al., 2005. Three-dimensional culture of newborn rat utricle using an extracellular matrix promotes formation of a cyst. Neuroscience, 133(1):253-265.

[17]Géléoc GS, Holt JR, 2014. Sound strategies for hearing restoration. Science, 344(6184):1241062.

[18]Gubb D, García-Bellido A, 1982. A genetic analysis of the determination of cuticular polarity during development in Drosophila melanogaster. J Embryol Exp Morphol, 68(1):37-57.

[19]Hawkins JE Jr, Johnsson LG, Aran JM, 1969. Comparative tests of gentamicin ototoxicity. J Infect Dis, 119(4-5):417-426.

[20]Hudspeth AJ, 1989a. How the ear’s works work. Nature, 341(6241):397-404.

[21]Hudspeth AJ, 1989b. Mechanoelectrical transduction by hair cells of the bullfrog’s sacculus. Prog Brain Res, 80: 129-135.

[22]Hudspeth AJ, 1997. How hearing happens. Neuron, 19(5):947-950.

[23]Kalinec GM, Webster P, Lim DJ, et al., 2003. A cochlear cell line as an in vitro system for drug ototoxicity screening. Audiol Neurootol, 8(4):177-189.

[24]Koehler KR, Hashino E, 2014. 3D mouse embryonic stem cell culture for generating inner ear organoids. Nat Protoc, 9(6):1229-1244.

[25]Landegger LD, Dilwali S, Stankovic KM, 2017. Neonatal murine cochlear explant technique as an in vitro screening tool in hearing research. J Vis Exp, (124):e55704.

[26]Lelli A, Asai Y, Forge A, et al., 2009. Tonotopic gradient in the developmental acquisition of sensory transduction in outer hair cells of the mouse cochlea. J Neurophysiol, 101(6):2961-2973.

[27]Li-Korotky HS, 2012. Age-related hearing loss: quality of care for quality of life. Gerontologist, 52(2):265-271.

[28]Lin JC, Zhang XD, Wu FF, et al., 2015. Hair cell damage recruited Lgr5-expressing cells are hair cell progenitors in neonatal mouse utricle. Front Cell Neurosci, 9:113.

[29]Malgrange B, Thiry M, van de Water TR, et al., 2002. Epithelial supporting cells can differentiate into outer hair cells and Deiters’ cells in the cultured organ of Corti. Cell Mol Life Sci, 59(10):1744-1757.

[30]Matz G, Rybak L, Roland PS, et al., 2004. Ototoxicity of ototopical antibiotic drops in humans. Otolaryngol-Head Neck Surg, 130(S3):S79-S82.

[31]May LA, Kramarenko II, Brandon CS, et al., 2013. Inner ear supporting cells protect hair cells by secreting HSP70. J Clin Invest, 123(8):3577-3587.

[32]May-Simera H, 2016. Evaluation of planar-cell-polarity phenotypes in ciliopathy mouse mutant cochlea. J Vis Exp, (108):e53559.

[33]Meiteles LZ, Raphael Y, 1994. Scar formation in the vestibular sensory epithelium after aminoglycoside toxicity. Hear Res, 79(1-2):26-38.

[34]Meyer-Bisch C, 2005. Measuring noise. Med Sci (Paris), 21(5):546-550.

[35]Oesterle EC, Rubel EW, 1993. Postnatal production of supporting cells in the chick cochlea. Hear Res, 66(2):213-224.

[36]Oesterle EC, Tsue TT, Reh TA, et al., 1993. Hair-cell regeneration in organ cultures of the postnatal chicken inner ear. Hear Res, 70(1):85-108.

[37]Ou HC, Lin V, Rubel EW, 2013. “In-bone” utricle cultures— a simplified, atraumatic technique for in situ cultures of the adult mouse (Mus musculus) utricle. Otol Neurotol, 34(2):353-359.

[38]Parker M, Brugeaud A, Edge AS, 2010. Primary culture and plasmid electroporation of the murine organ of Corti. J Vis Exp, (36):e1685.

[39]Qian D, Jones C, Rzadzinska A, et al., 2007. Wnt5a functions in planar cell polarity regulation in mice. Dev Biol, 306(1):121-133.

[40]Quint E, Hackney CM, Furness DN, 1996. The effect of neomycin on organotypic cultures of the adult guinea-pig utricle. Ann N Y Acad Sci, 781(1):683-685.

[41]Rastel D, Abdouh A, Dahl D, et al., 1993. An original organotypic culture method to study the organ of Corti of the newborn rat in vitro. J Neurosci Methods, 47(1-2):123-131.

[42]Roberson DW, Rubel EW, 1994. Cell division in the gerbil cochlea after acoustic trauma. Am J Otol, 15(1):28-34.

[43]Romand R, Chardin S, 1999. Effects of growth factors on the hair cells after ototoxic treatment of the neonatal mammalian cochlea in vitro. Brain Res, 825(1-2):46-58.

[44]Ruben RJ, 1967. Development of the inner ear of the mouse: a radioautographic study of terminal mitoses. Acta Otolaryngol, Suppl 220:1-44.

[45]Shang JL, Cafaro J, Nehmer R, et al., 2010. Supporting cell division is not required for regeneration of auditory hair cells after ototoxic injury in vitro. J Assoc Res Otolaryngol, 11(2):203-222.

[46]Smith ME, Groves AK, Coffin AB, 2016. Editorial: sensory hair cell death and regeneration. Front Cell Neurosci, 10:208.

[47]Sobkowicz HM, Bereman B, Rose JE, 1975. Organotypic development of the organ of Corti in culture. J Neurocytol, 4(5):543-572.

[48]Spencer NJ, Cotanche DA, Klapperich CM, 2008. Peptide- and collagen-based hydrogel substrates for in vitro culture of chick cochleae. Biomaterials, 29(8):1028-1042.

[49]Taura A, Nakashima N, Ohnishi H, et al., 2016. Regenerative therapy for vestibular disorders using human induced pluripotent stem cells (iPSCs):neural differentiation of human iPSC-derived neural stem cells after in vitro transplantation into mouse vestibular epithelia. Acta Otolaryngol, 136(10):999-1005.

[50]von Békésy G, 1956. Current status of theories of hearing. Science, 123(3201):779-783.

[51]Werner M, van de Water TR, Andersson T, et al., 2012. Morphological and morphometric characteristics of vestibular hair cells and support cells in long term cultures of rat utricle explants. Hear Res, 283(1-3):107-116.

[52]Werner M, van de Water TR, Hammarsten P, et al., 2015. Morphological and morphometric characterization of direct transdifferentiation of support cells into hair cells in ototoxin-exposed neonatal utricular explants. Hear Res, 321:1-11.

[53]White PM, Doetzlhofer A, Lee YS, et al., 2006. Mammalian cochlear supporting cells can divide and trans-differentiate into hair cells. Nature, 441(7096):984-987.

[54]Yamahara K, Yamamoto N, Nakagawa T, et al., 2015. Insulin-like growth factor 1: a novel treatment for the protection or regeneration of cochlear hair cells. Hear Res, 330:2-9.

[55]Yamashita T, Vosteen KH, 1975. Tissue culture of the organ of Corti and the isolated hair cells from the newborn guinea pig. Acta Otolaryngol, 79(S330):77-90.

[56]Zhao HB, 2001. Long-term natural culture of cochlear sensory epithelia of guinea pigs. Neurosci Lett, 315(1-2):73-76.