Predegenerated Schwann cells promote neuroprotection and regeneration of retinal ganglion cells in ex vivo rat retinal explants

Abstract

AbstractPurpose To investigate neuroprotective and pro‐regenerative potential of predegenerated Schwann cells towards RGCs in ex vivo retinal explants.Methods In this study we used two models of rat retinal explants culture ‐ reversed explants from P2 Wistar rats pups and insert explants from 4 weeks old Wistar rats. Both types of explants were cultured in Neurobasal A medium with supplements (B‐27, N2 and GlutaMax). In this way we prepared 32 insert explants (16 were cultured with 2 µl of Schwann cells suspension (104 cells) dropped on the each explants surface, another 16 served as control treated with 2 µl of DPBS) and 30 reversed explants (10 of them were cultured in standard medium, 10 explants were with addition of 5 ng/ml BDNF into medium and 10 were co‐cultured with 104 of Schwann cells). After proper culture time, explants were fixed and processed for immunostainings and stereological analyses.Results In retinal explants cultured in inserts system, there was significantly higher density of RGCs in group treated with Schwann cells in comparison to PBS treated (independent samples t‐test, p<0.05). In reversed retinal explants, there were significant differences between groups expressed in mean and total length and surface of neurites, as well as in number and arboration rate of neurites (U‐Mann Whitney test, p<0.05). Immunofluorescent staining using GAP43 additionally revealed intense neurites outgrowth (also in explants from adult animals).Conclusion Predegenerated Schwann cells exert neuroprotective action and support regeneration of RGCs in neonatal and adult retina in ex vivo conditions.