Astrocytes Regulate the Development and Maturation of Retinal Ganglion Cells Derived from Human Pluripotent Stem Cells

Abstract

SummaryRetinal ganglion cells (RGCs) form the connection between the eye and the brain, with this connectivity disrupted in numerous blinding disorders. Previous studies have demonstrated the ability to derive RGCs from human pluripotent stem cells (hPSCs); however, these cells exhibited some characteristics that indicated a limited state of maturation. Among the many factors known to influence RGC development in the retina, astrocytes are known to play a significant role in their functional maturation. Thus, efforts of the current study examined the functional maturation of hPSC-derived RGCs, including the ability of astrocytes to modulate this developmental timeline. Morphological and functional properties of RGCs were found to increase over time, with astrocytes significantly accelerating the functional maturation of hPSC-derived RGCs. The results of this study clearly demonstrate the functional and morphological maturation of RGCs in vitro, including the effects of astrocytes on the maturation of hPSC-derived RGCs.

Plasma Rich in Growth Factors Enhances Cell Survival after in Situ Retinal Degeneration

“…This tissue is composed of different cell types, such as neuronal cells, photoreceptors, and epithelial cells, which are involved in the biochemical processes that are carried out in this tissue. For instance, photoreceptors oversee the process of sending electrical stimuli through the optical nerve to the brain to create images [ 36 , 37 ]. Epithelial cells are involved in the protection of and nutrient transport to photoreceptors [ 38 ].…”

Retinal ganglion cells harboring the OPTN(E50K) mutation exhibit neurodegenerative phenotypes when derived from hPSC-derived three dimensional retinal organoids

“…In this capacity, several disease-related phenotypes were demonstrated to be specific to RGCs compared to other retinal cell types. However, as RGCs are the projection neurons of the retina that extend long and intricate neurites to transmit visual information to the brain, the morphological maturation and functionality of RGCs is sometimes easier to interpret when these cells are isolated from retinal organoids and grown in 2D culture (VanderWall et al, 2019). Results of the current study took advantage of both 3D organoid approaches as well as 2D RGC cultures to understand RGC development and maturation as well as identify disease phenotypes in these conditions.…”

“…Using previously described protocols, single cell suspensions were then enriched for RGCs with the Thy1.2 surface receptor using the MACS magnetic cell separation kit (Sluch et al, 2017). 10,000 RGCs were plated on poly-d-ornithine and laminin-coated 12 mm coverslips and maintained for up to 4 weeks in BrainPhys Media (VanderWall et al, 2019). To analyze neurite complexity, RGCs were transfected with GFP using lipofectamine 2 days before fixation to aid in identification of individual RGC neurites along with BRN3B:tdTomato expression.…”

“…Whole-cell patch clamp recordings were performed at room temperature (∼22 °C) using a HEKA EPC-10 amplifier as previously described (VanderWall et al, 2019). The PatchMaster program was used for data acquisition.…”

Improved Ocular Tissue Models and Eye-On-A-Chip Technologies Will Facilitate Ophthalmic Drug Development

“…Ultimately, to be used for drug screening, both the organoid and tissue chip fields need to overcome the challenges of Human pluripotent stem cell-derived astrocytes enhance maturation of retinal ganglion cells 28…”