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Method Article
Natalie Bull
Keith Martin University of Cambridge (UK)
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol details a method for isolating retinal tissue from adult rats as an organotypic culture to study neurobiological processes in mature tissue. It combines the efficiency and control common to in vitro techniques with close imitation of the in vivo environment. Eyes from adult rats are enucleated and the neural retina is isolated. Tissue is cut into quarters, yielding eight retinal explants per animal, and cultured at a fluid/air interface on organotypic culture membranes. Explantation can be accomplished in thirty minutes per animal. Tissue is nourished by a serum-free medium and can be viably maintained for at least two weeks ex vivo. Protocols are provided that describe histological processing, including techniques for whole-mount and cross-sectional immunohistochemical labelling. In addition, methods for simulating intraocular cell transplantation and pharmacological screening for neuroprotective therapies are also provided.
Keywords
neuroprotection, ganglion cell, organotypic culture, pharmacological screening
The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.pdf
Retinal Explant Protocol PDF Complete PDF of Martin Lab Retinal Explant Protocol
- supplement0.pdf
Reagents Reagents
- supplement0.pdf
Troubleshooting Troubleshooting
- supplement0.pdf
Equipment Equipment
- supplement0.pdf
Introduction Introduction
- supplement0.pdf
Timing Timing
- supplement0.pdf
Anticipated Results Anticipated Results
- supplement0.pdf
References References
- supplement0.pdf
Acknowledgements Acknowledgements
- supplement0.pdf
Procedure Procedure
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Associated Publications
Use of an adult retinal explant model for screening of potential retinal ganglion cell neuroprotective therapies
by N. D. Bull, T. V. Johnson, G. Welsapar, +3
Investigative Ophthalmology & Visual Science (02 March, 2011)
Development and Characterization of an Adult Retinal Explant Organotypic Tissue Culture System as an In Vitro Intraocular Stem Cell Transplantation Model
by T. V. Johnson and K. R. Martin
Investigative Ophthalmology & Visual Science (02 March, 2011)
Identification of Barriers to Retinal Engraftment of Transplanted Stem Cells
by T. V. Johnson, N. D. Bull, and K. R. Martin
Investigative Ophthalmology & Visual Science (02 March, 2011)
Method Article
Natalie Bull
Keith Martin University of Cambridge (UK)
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 04 Mar, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
This protocol details a method for isolating retinal tissue from adult rats as an organotypic culture to study neurobiological processes in mature tissue. It combines the efficiency and control common to in vitro techniques with close imitation of the in vivo environment. Eyes from adult rats are enucleated and the neural retina is isolated. Tissue is cut into quarters, yielding eight retinal explants per animal, and cultured at a fluid/air interface on organotypic culture membranes. Explantation can be accomplished in thirty minutes per animal. Tissue is nourished by a serum-free medium and can be viably maintained for at least two weeks ex vivo. Protocols are provided that describe histological processing, including techniques for whole-mount and cross-sectional immunohistochemical labelling. In addition, methods for simulating intraocular cell transplantation and pharmacological screening for neuroprotective therapies are also provided.
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Associated Publications
Use of an adult retinal explant model for screening of potential retinal ganglion cell neuroprotective therapies
by N. D. Bull, T. V. Johnson, G. Welsapar, +3
Investigative Ophthalmology & Visual Science (02 March, 2011)
Development and Characterization of an Adult Retinal Explant Organotypic Tissue Culture System as an In Vitro Intraocular Stem Cell Transplantation Model
by T. V. Johnson and K. R. Martin
Investigative Ophthalmology & Visual Science (02 March, 2011)
Identification of Barriers to Retinal Engraftment of Transplanted Stem Cells
by T. V. Johnson, N. D. Bull, and K. R. Martin
Investigative Ophthalmology & Visual Science (02 March, 2011)