Scientific Publication: Coculture of Axotomized Rat Retinal Ganglion Neurons with Olfactory Ensheathing Glia, as an In Vitro Model of Adult Axonal Regeneration. Portela-Lomba et al., Journal of Visualized Experiments (JoVE) 2020
Professor M. Teresa Moreno-Flores, in collaboration with the researchers of the Universidad Francisco de Vitoria María Portela-Lomba (predoctoral), Diana Simón y Javier Sierra, and of the Universidad de Catania, Cristina Russo, have published this work in the international Journal of Visualized Experiments (JoVE). The publication describes and visually illustrates the use of a technique to evaluate and quantify in vitro the capacity of olfactory ensheathing glia cells to foster adult CNS neurons axonal regeneration.
Olfactory ensheathing glia (OEG) cells are localized all the way from the olfactory mucosa to and into the olfactory nerve layer (ONL) of the olfactory bulb. Throughout adult life, they are key for axonal growing of newly generated olfactory neurons, from the lamina propria to the ONL. Due to their pro-regenerative properties, these cells have been used to foster axonal regeneration in spinal cord or optic nerve injury models. We present an in vitro model to assay and measure OEG neuroregenerative capacity after neural injury. In this model, reversibly immortalized human OEG (ihOEG) is cultured as a monolayer, retinas are extracted from adult rats and retinal ganglion neurons (RGN) are cocultured onto the OEG monolayer. After 96 h, axonal and somatodendritic markers in RGNs are analyzed by immunofluorescence and the number of RGNs with axon and the mean axonal length/neuron are quantified. This protocol has the advantage over other in vitro assays that rely on embryonic or postnatal neurons, that it evaluates OEG neuroregenerative properties in adult tissue. In addition, it is not only useful for assessing the neuroregenerative potential of ihOEG but can be extended to different sources of OEG or other glial cells.
Portela-Lomba M, Simón D, Russo C, Sierra J, Moreno-Flores MT (2020). Coculture of Axotomized Rat Retinal Ganglion Neurons with Olfactory Ensheathing Glia, as an In Vitro Model of Adult Axonal Regeneration. J Vis Exp. 2020 Nov 2;(165). doi: 10.3791/61863.
https://www.jove.com/t/61863/coculture-axotomized-rat-retinal-ganglion-neurons-with-olfactory