Journal
STEM CELLS
Volume 37, Issue 5, Pages 593-598Publisher
OXFORD UNIV PRESS
DOI: 10.1002/stem.2963
Keywords
Pluripotent stem cells; Single cell RNA-Seq; Retinal organoids
Categories
Funding
- ERC [614620]
- RP Fighting Blindness [GR593]
- BBSRC [BB/I02333X/1]
- MRC [MR/M008886/1, MC_PC_15030]
- European Research Council (ERC) [614620] Funding Source: European Research Council (ERC)
- BBSRC [BB/I02333X/1] Funding Source: UKRI
- MRC [MC_UU_00015/9, MR/M008886/1, MC_PC_15030] Funding Source: UKRI
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The rapid improvements in single cell sequencing technologies and analyses afford greater scope for dissecting organoid cultures composed of multiple cell types and create an opportunity to interrogate these models to understand tissue biology, cellular behavior and interactions. To this end, retinal organoids generated from human embryonic stem cells (hESCs) were analyzed by single cell RNA-sequencing (scRNA-Seq) at three time points of differentiation. Combinatorial data from all time points revealed the presence of nine clusters, five of which corresponded to key retinal cell types: retinal pigment epithelium (RPE), retinal ganglion cells (RGCs), cone and rod photoreceptors, and Muller glia. The remaining four clusters expressed genes typical of mitotic cells, extracellular matrix components and those involved in homeostasis. The cell clustering analysis revealed the decreasing presence of mitotic cells and RGCs, formation of a distinct RPE cluster, the emergence of cone and rod photoreceptors from photoreceptor precursors, and an increasing number of Muller glia cells over time. Pseudo-time analysis resembled the order of cell birth during retinal development, with the mitotic cluster commencing the trajectory and the large majority of Muller glia completing the time line. Together, these data demonstrate the feasibility and potential of scRNA-Seq to dissect the inherent complexity of retinal organoids and the orderly birth of key retinal cell types.
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