hPSC-derived Photoreceptor Synaptic Puncta and Synaptic Tracing Datasets and Code

Data used for analysis of synaptic puncta formation and synaptic tracing of human pluripotent stem cell (hPSC)-derived photoreceptors (PRs). This data is deposited as supplemental material for a manuscript by Allison L. Ludwig, Steven J. Mayerl, Yu Gao, Mark Banghart, Cole Bacig, Maria A. Fernandez Zepeda, Xinyu Zhao, and David M. Gamm, entitled “Re-formation of synaptic connectivity in dissociated human stem cell-derived retinal organoid cultures.” Please see associated manuscript for further details. Summary: These data are the result of manual and automated cell counts from confocal and epifluorescence microscopy experiments using dissociated hPSC-PRs differentiated from the WA09 wild type stem cell line to explore synaptic re-connection following dissociation of early retinal organoids. The included files consist of raw cell count data (.csv files) and R code (.Rmd files) used for analysis of synaptic puncta formation and synaptic tracing of hPSC-PRs in the days following dissociation from retinal organoids. The results of these experiments demonstrate detection of synaptic puncta within hPSC-PRs as early as 2 days after dissociation, with a stepwise increase in the proportion of hPSC-PRs with organized synaptic puncta on days 5 and 20 post-dissociation. Moreover, we observed consistent and measurable re-formation of synaptic connections after dissociation from retinal organoids, as demonstrated by monosynaptic retrograde rabies viral tracing, even after controlling for biomaterial transfer. These results demonstrate plasticity of hPSC-PRs and further underscores their potential to restore visual circuits.