title: Intercellular Adhesion-Dependent Cell Survival and ROCK-Regulated Actomyosin-Driven Forces Mediate Self-Formation of a Retinal Organoid.
abstract: In this study we dissected retinal organoid morphogenesis in human embryonic stem cell (hESC)-derived cultures and established a convenient method for isolating large quantities of retinal organoids for modeling human retinal development and disease. Epithelialized cysts were generated via floating culture of clumps of Matrigel/hESCs. Upon spontaneous attachment and spreading of the cysts, patterned retinal monolayers with tight junctions formed. Dispase-mediated detachment of the monolayers and subsequent floating culture led to self-formation of retinal organoids comprising patterned neuroretina, ciliary margin, and retinal pigment epithelium. Intercellular adhesion-dependent cell survival and ROCK-regulated actomyosin-driven forces are required for the self-organization. Our data supports a hypothesis that newly specified neuroretina progenitors form characteristic structures in equilibrium through minimization of cell surface tension. In long-term culture, the retinal organoids autonomously generated stratified retinal tissues, including photoreceptors with ultrastructure of outer segments. Our system requires minimal manual manipulation, has been validated in two lines of human pluripotent stem cells, and provides insight into optic cup invagination in vivo.
authors.affiliation: Department of Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Albert
authors.initials: A
authors.lastname: Lowe
authors.affiliation: Department of Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Raven
authors.initials: R
authors.lastname: Harris
authors.affiliation: Department of Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Punita
authors.initials: P
authors.lastname: Bhansali
authors.affiliation: Department of Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Department of Ophthalmology and Visual Sciences, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Ales
authors.initials: A
authors.lastname: Cvekl
authors.affiliation: Department of Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA; Department of Ophthalmology and Visual Sciences, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Electronic address: wei.liu@einstein.yu.edu.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
mesh_terms: D000205:Actomyosin / Q000235:genetics; D002448:Cell Adhesion / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics*; D002470:Cell Survival / Q000235:genetics; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D009024:Morphogenesis / Q000235:genetics; D009940:Organoids / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D012160:Retina / Q000254:growth & development / Q000378:metabolism*; D055213:Retinal Pigment Epithelium / Q000254:growth & development / Q000378:metabolism; D054460:rho-Associated Kinases / Q000235:genetics*
chemical_list: D000205:Actomyosin; D054460:rho-Associated Kinases
keywords: Rho kinase (ROCK); actomyosin; apicobasal cell polarity; cell adhesion; cell survival; cell-surface tension; differentiation; epithelialization; extracellular matrix; human embryonic stem cells; retinal progenitor cells; self-organizing organoids