Single cell transcriptomic comparison of human fetal retina, hPSC-derived retinal organoids, and long-term retinal cultures.

To study the development of human retina, we used single cell RNAseq at key fetal stages and followed the development of the major cell types, as well as populations of transitional cells. We also analyzed stem cell (hPSC)-derived retinal organoids; although organoids have a very similar cellular composition at equivalent ages to the fetal retina, there are some differences in gene expression of particular cell types. Moreover, the inner retinal lamination is disrupted in more advanced stages of organoids when compared with fetal retina. To determine whether the disorganization in the inner retina was due to the culture conditions, we analyzed retinal development in fetal retina maintained under similar conditions. These retinospheres develop for at least 6 months, displaying better inner retinal lamination than retinal organoids. Our scRNAseq comparisons between fetal retina, retinal organoids and retinospheres provide a new resource for developing better in vitro models for retinal disease.

为研究人类视网膜的发育进程，我们采用关键胎儿发育阶段的单细胞RNA测序（single cell RNAseq）技术，对主要细胞类型以及过渡细胞群的发育过程进行了追踪。我们还分析了由人多能干细胞（human pluripotent stem cell, hPSC）诱导分化得到的视网膜类器官；尽管在对应发育阶段，类器官的细胞组成与胎儿视网膜高度相似，但特定细胞类型的基因表达仍存在一定差异。此外，相较于胎儿视网膜，发育至更晚期阶段的类器官其视网膜内层分层结构会出现紊乱。为探究视网膜内层结构紊乱是否由培养条件所导致，我们对置于相同培养环境下的胎儿视网膜进行了体外培养分析。这些视网膜球（retinospheres）可在体外维持发育至少6个月，其视网膜内层分层结构相较于视网膜类器官更为完整。我们针对胎儿视网膜、视网膜类器官及视网膜球开展的单细胞RNA测序对比分析，可为构建更优化的视网膜疾病体外模型提供全新的研究资源。