A novel method for co-culture with Müller cells and microglia in rat retina in vitro.

Microglia and Müller cells are glial cells of the retina and constitute a functional link between neurons and vessels. The aim of the present study was to introduce a novel method of co-culture with Müller cells and microglia in rat retina. A camera was used to analyze all the cell changes. Immunofluorescence staining of glutamine synthetase and OX-42 were used for the identification of Müller cells and microglial, respectively. On day 1, all the cell types from the retina were round or oval and floating in the medium. On the following days, microglial cells were adherent and proliferated. Müller cells stretched and quickly proliferated. On days 12-15, microglial cells were floating in the medium. Following agitation, microglial cells became quickly detached from the flask walls, whereas Müller cells remained adherent. In conclusion, agitation is an effective way to separate microglial cells from Müller cells. The time of detachment and the speed of agitation are essential. Co-culture with Müller cells and microglia in the retina is economical and useful for future methods in microglia and Müller cell research.