In vitro characterisation of spontaneously immortalised Non-Human Primate Müller glia cell lines as a potential source for cell replacement therapies for retinal degenerative eye disease

Abstract

Purpose : Müller glia, which are astrocyte-like radial cells that play a pivotal role in maintaining retinal homeostasis, are considered to be the main glial cells in the retina. Their expression spans along the retina providing general structural support to retinal neurones and blood vessels. Among several other functions, they prevent aberrant photoreceptors migration into the subretinal space and facilitate glutamate uptake to keep its extracellular concentration below toxic levels.
In lower vertebrates such as fish and amphibians, the injured retina shows a regeneration potential which is believed to stem from Müller glia. Although this regeneration potential is common for all vertebrates, it is, however, absent in mammalian systems for unknown reasons. Although various Müller glia cell lines have been described in the literature including those derived from human and rat, to our knowledge, no non-human primate (NHP) Müller glia cell line is currently available.

Methods : A spontaneously immortalised Müller glia cell line was established from a primary culture of neural retina of a Macaque monkey dissociated using a papain dissociation reagent. The cells were cultured, passaged and eventually immortalised without the presence of growth factors. The cells were examined for expression of Müller glia and other markers by immunocytochemistry and RT-PCR and differentiated with growth factors known to stimulate cells differentiation.

Results : Here we report a spontaneously immortalised Müller glia cell line isolated from rhesus macaque monkeys, grown under normal culture conditions and could be expanded indefinitely without the presence of growth factors. The cell line exhibits normal morphology and expressed Müller glia markers among other stem cell markers when examined with immunocytochemistry and RT-PCR. When cultured in the presence of growth factors that stimulate cell differentiation, they altered their gene expression profile by expressing a combination of retinal neuronal markers

Conclusions : Our observations indicate that the non-human primate retina harbours a population of cells that express both Müller glia and stem cell markers, which can be useful for future cell replacement therapies.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.