Open AccessDeconstructing Retinal Organoids: Single Cell RNA‐Seq Reveals the Cellular Components of Human Pluripotent Stem Cell‐Derived Retina
Author(s) -
Collin Joseph,
Queen Rachel,
Zerti Darin,
Dorgau Birthe,
Hussain Rafiqul,
Coxhead Jonathan,
Cockell Simon,
Lako Majlinda
Publication year - 2019
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.2963
Subject(s) - biology , microbiology and biotechnology , embryonic stem cell , stem cell , retina , retinal pigment epithelium , induced pluripotent stem cell , retinal , organoid , muller glia , cell type , cell , cellular differentiation , progenitor cell , genetics , neuroscience , gene , botany
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 Müller 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 Müller 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 Müller 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. S tem C ells 2019;37:593–598