Single cell sequencing of whole eye organoids to identify ocular cell susceptible to infection with SARS-CoV2 and the viral response in infected cells

We diffrenciated organoid cultures reprecenting ocular cell, from human pluipotent stem cells (hPSCs). The organoids were grown following a protocol for self-formed ectodermal autonomous multizone (SEAM) where the different orcular cells will form in concentric circles (zone) representing surface ectoderm, cornea, RPE, neural progenitors, dorsal optic cup and periocular mesenchyme. We use this eye organoid to investigate whether the ocular surface is sucessable to infection with SARC-CoV-2. We show that a sup-group of cells get readerly infected after expositor to the virus through the media, at 1MOI for 24hours. We identify this cells population as limbal/corneal cells. By anchoring the data from the infected SEAM culture to a not-infected SEAM culture we investigated the cellular response to infection. We find that both a NFkB and a interferon response is mounted, and interestingly the Interferon pathway genes are suppressed in the infected cells compared to the nearest neighbor clusters while the NFkB pathway is more highly expressed in the infected cell compared to neighbor cells. Single cell sequencing of a SEAM culture infected with SARS-CoV2 by exposure to the virus through the media at 1MOI for 24h and not-infected SEAM culture.