Advertisement

Three-Dimensional Culture of Mouse Eyecups

  • Raven Diacou
  • Punita Bhansali
  • Wei LiuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2092)

Abstract

Retinal neurons and glia in the adult vertebrate retina are differentiated from multipotent retinal progenitors in the eyecups under the regulation of intrinsic and extrinsic factors, but the molecular mechanism underlying the process is partially understood. Functional studies using engineered mice provide tremendous insight into the mechanisms of retinal cell differentiation, but in utero embryogenesis prevents manipulations of mouse embryonic retina. Mouse eyecup culture using a culture filter or insert has been developed, but retinal structure is often altered due to the flattening of mouse eyecups in these culture systems. In this chapter, we describe three-dimensional culture of embryonic mouse eyecups. In our system, cell differentiation, stratified retinal structure, and ciliary margins in cultured eyecups were reminiscent of those in vivo. Our 3D culture of mouse eyecups has multiple applications when wild-type or engineered mice are used as models for studying retinal cell differentiation.

Key words

Retinal cell differentiation Three-dimensional culture Mouse eyecup explant 

Notes

Acknowledgments

We are thankful to Dr. R. Chuck for the support, grants from BrightFocus (M2012044 to W.L.), NEI (1R01EY022645 to W.L.), and RPB (unrestricted grant to the Department of Ophthalmology and Visual Science at Albert Einstein College of Medicine). R.H. and P.B. were supported by NIH training grants T32GM007288 and K12GM102779, respectively.

References

  1. 1.
    Cepko C (2014) Intrinsically different retinal progenitor cells produce specific types of progeny. Nat Rev Neurosci 15(9):615–627.  https://doi.org/10.1038/nrn3767CrossRefPubMedGoogle Scholar
  2. 2.
    Donovan SL, Dyer MA (2006) Preparation and square wave electroporation of retinal explant cultures. Nat Protoc 1(6):2710–2718.  https://doi.org/10.1038/nprot.2006.454CrossRefPubMedGoogle Scholar
  3. 3.
    Jin K, Xiang M (2012) In vitro explant culture and related protocols for the study of mouse retinal development. Methods Mol Biol 884:155–165.  https://doi.org/10.1007/978-1-61779-848-1_10CrossRefPubMedGoogle Scholar
  4. 4.
    Liu H, Xu S, Wang Y, Mazerolle C, Thurig S, Coles BL, Ren JC, Taketo MM, van der Kooy D, Wallace VA (2007) Ciliary margin transdifferentiation from neural retina is controlled by canonical Wnt signaling. Dev Biol 308(1):54–67.  https://doi.org/10.1016/j.ydbio.2007.04.052CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Ophthalmology and Visual SciencesAlbert Einstein College of MedicineBronxUSA
  2. 2.Department of GeneticsAlbert Einstein College of MedicineBronxUSA
  3. 3.Department of ​Biological Sciences & GeologyQueensborough Community CollegeBaysideUSA

Personalised recommendations