Mouse retinal explants
Scope of the method
- Animal health
- Basic Research
- In vitro - Ex vivo
- Animal derived cells / tissues / organs
Description
- Tissue explant
- retina
- neurite outgrowth
- immunohistochemical staining
- automated morphometric analysis
- preserving cell-to-cell interaction
- axonal regeneration
- neurodegeneration
- retinal differentation
Organotypical culture models, such as retinal explants, is an ideal alternative for in vitro retinal cell cultures and preclinical animal models, as they provide the necessary compromise between these two model systems. The major advantage of explant cultures is that cells are kept within their normal environment, thereby preserving cell-to-cell interactions, while maintaining a higher level of experimental control as in animal models. Organotypic cultures thereby provide an ideal platform for the identification and validation of novel neuroprotective or pro-regenerative substances. As retinal explants have previously been used to study various neural processes such as neurodegeneration and neurite outgrowth, they provide an ideal ex vivo system to screen promising molecules in an in vivo-like situation. This technique includes retinal explant dissection, culture, immunostaining, and automated analysis methods using ImageJ.
- Dissection microscope with light source ;
- Horizontal laminar flow ;
- Dissection material ;
- Incubator ;
- Confocal microscope.
- Internally validated
- Published in peer reviewed journal
Pros, cons & Future potential
The major advantage of explant cultures is that cells are kept within their normal environment, thereby preserving cell-to-cell interactions. They provide an ideal ex vivo system to screen promising molecules.
No time-lapse experiments no objective distinction can be made between glial and neuronal processes, which may result in a false representation of neurite outgrowth.
Transgenic animals, that express a fluorescent protein in RGC axons, such as the thy1-YFP mice might allow for time-lapse experiments.
- In diabetic retinopathy, the retinal explants will be used to test compounds known to be involved in developing perivascular membranes.
- Ideal technique for screening potential regenerative molecules, both in mouse and zebrafish models.
References, associated documents and other information
BOLLAERTS, I., VAN HOUCKE, J., ANDRIES, L., DE GROEF, L. & MOONS, L. 2017. Neuroinflammation as Fuel for Axonal Regeneration in the Injured Vertebrate Central Nervous System. Mediators Inflamm, 2017, 9478542.
BUYENS, T., GAUBLOMME, D., VAN HOVE, I., DE GROEF, L. & MOONS, L. 2014. Quantitative assessment of neurite outgrowth in mouse retinal explants. Methods Mol Biol, 1162, 57-71. GAUBLOMME, D., BUYENS, T. & MOONS, L. 2013. Automated analysis of neurite outgrowth in mouse retinal explants. J Biomol Screen, 18, 534-43.
Contact person
Evy LefevereOrganisations
Katholieke Universiteit Leuven (KUL)Biology
Belgium
Flemish Region