Small molecules DAPT and Forskolin enhances ganglion cells expression in porcine retinal progenitor cells

Abstract

Abstract Background Retinal ganglion cell (RGCs) loss underlies several retinal degenerative diseases, including Glaucoma and NF1-OPG. RGCs are essential cells in the visual pathway. These cells transmit the signal gathered from the retinal photoreceptors to the brain via the optic nerve and loss of these cells leads to permanent loss of vision. Presently there is no treatment, however, in the last decade cell replacement has shown potential to restore vision in these conditions. Materials and Methods In this study, we have established protocol with a dose dependent response differentiating porcine retinal progenitors (pRPCs) into retinal ganglion cells (RGC) with DAPT and Forskolin treatments. Cells were initially treated with a known concentration of DAPT and Forskolin that has been widely used to culture RGCs. Results We identify the optimal concentration and time course that yields the highest expression of RGCs, pRPCs were cultured for 4 weeks in 25 different concentrations of small molecules. At week 2 and 3 approximately 40% of the pRPCs population expressed the neuronal marker TUJ1 and RGCs markers such as RBPMS and Thy1.1. Differentiated cells showed the lowest proliferative index as indicated by Ki-67 expression. Conclusion This data demonstrates that specific concentrations of DAPT and Forskolin can induce RPCs to undergo differentiation into RGCs linage.