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Organotypic Culture of the Mouse Cochlea from Embryonic Day 12 to the Neonate

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Auditory and Vestibular Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1427))

Abstract

The development of the mammalian cochlea is a complex process involving several intersecting signaling pathways to ultimately generate its highly organized cellular architecture. In humans, and in the mouse, there is one row of inner hair cells aligned next to three rows of outer hair cells. The support cells intercalate between the hair cells to create a cellular mosaic across the organ of Corti (OC). Organotypic culture of the cochlea is a valuable technique for investigating the early stages of OC development. Cultures can be established at proliferative stages and maintained in vitro until cellular differentiation commences. It is straightforward to monitor differentiation in response to perturbations of key signaling pathways using pharmacological agents. While postnatal cochlea organ cultures have already been adapted for in vitro studies, it is more challenging to establish cultures at early embryonic stages due to the small size of the organ primordium. The protocol described in this chapter is permissive for all stages of development from E12 cochleas to day 5 neonatal murine cochleas, allowing culture survival up to 2 weeks in vitro.

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References

  1. Oumard A, Qiao J, Jostock T, Li J, Bode J (2006) Recommended method for chromosome exploitation: RMCE-based cassette-exchange systems in animal cell biotechnology. Cytotechnology 50(1–3):93–108. doi:10.1007/s10616-006-6550-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Brigande JV, Gubbels SP, Woessner DW, Jungwirth JJ, Bresee CS (2009) Electroporation-mediated gene transfer to the developing mouse inner ear. Methods Mol Biol 493:25–139. doi:10.1007/978-1-59745-523-7_8

    Google Scholar 

  3. Gubbels SP, Woessner DW, Mitchell JC, Ricci AJ, Brigande JV (2008) Functional auditory hair cells produced in the mammalian cochlea by in utero gene transfer. Nature 455(7212):537–541. doi:10.1038/nature07265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wang L, Jiang H, Brigande JV (2012) Gene transfer to the developing mouse inner ear by in vivo electroporation. J Vis Exp (64). doi: 3791/3653

    Google Scholar 

  5. Munnamalai V, Hayashi T, Bermingham-McDonogh O (2012) Notch prosensory effects in the mammalian cochlea are partially mediated by Fgf20. J Neurosci 32(37):12876–12884. doi:10.1523/JNEUROSCI.2250-12.2012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Hayashi T, Kokubo H, Hartman BH, Ray CA, Reh TA, Bermingham-McDonogh O (2008) Hesr1 and Hesr2 may act as early effectors of Notch signaling in the developing cochlea. Dev Biol 316(1):87–99. doi:10.1016/j.ydbio.2008.01.006

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Chen SY, Hayashida Y, Chen MY, Xie HT, Tseng SC (2011) A new isolation method of human limbal progenitor cells by maintaining close association with their niche cells. Tissue Eng Part C Methods 17(5):537–548. doi:10.1089/ten.TEC.2010.0609

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Gong Q (2012) Culture of mouse olfactory sensory neurons. Curr Protoc Neurosci Chapter 3:Unit3 24. doi: 10.1002/0471142301.ns0324s58

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Acknowledgments

The authors thank the members of the Fekete lab for helpful suggestions included in the protocol. Work in the Fekete lab using this culture technique is funded by NIH/NIDCD grant R01 DC002756.

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Correspondence to Vidhya Munnamalai .

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© 2016 Springer Science+Business Media New York

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Munnamalai, V., Fekete, D.M. (2016). Organotypic Culture of the Mouse Cochlea from Embryonic Day 12 to the Neonate. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology, vol 1427. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3615-1_17

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  • DOI: https://doi.org/10.1007/978-1-4939-3615-1_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3613-7

  • Online ISBN: 978-1-4939-3615-1

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