Abstract
Purpose :
Equipping the human retinal organoids (hROs) with neuro-immune cells (retinal microglia) to make hROs more representative of real retinas.
Methods :
An efficient and simplified method was established to generate high-purity microglia from human pluripotent stem cells (hPSCs), then co-culturing the microglia with hROs to induce the differentiation of retinal microglia. The retinal microglia were characterized by the flow cytometry, ELISA, immunofluorescence staining and RNA sequencing.
Results :
We established a simplified approach to differentiate hPSCs into high purity (>90%) microglia (PSC-MG). PSC-MG express microglia-specific markers, release cytokines upon stimulation, and are capable of phagocytizing bacteria. When co-cultured with three-dimensional hROs, PSC-MG migrated into the hROs, tended to differentiate into resident retinal microglia, and simultaneously induced apoptosis in some neural cells and promoted the migration of the photoreceptor precursors.
Conclusions :
We developed a simplified and efficient method to generate microglia from human pluripotent stem cells, and we reported the first derivation of retina-resident microglia in vitro, providing a new source of human retinal microglia for developmental and disease studies and regenerative therapeutics.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.