Hydrogel-based milliwell arrays for standardized and scalable retinal organoid cultures

• Published in: Scientific reports 2020-06, Vol.10 (1), p.10275-10275, Article 10275
• Main Authors: Decembrini, S., Hoehnel, S., Brandenberg, N., Arsenijevic, Y., Lutolf, M. P.
• Format: Article
• Language: English
• Subjects: 13; 13/100; 13/107; 631/136/532/1360; 631/532/2117; 631/61/32; 639/166/985; 639/301/923/1027; 64; 82; 96; 96/100; 96/47; Aggregates; Animal models; Animals; Biomimetic Materials - chemistry; Biomimetics; Cell culture; Cell Culture Techniques - methods; Cell Differentiation; Cell Line; Cones; Embryo cells; Heterotrimeric GTP-Binding Proteins - analysis; Heterotrimeric GTP-Binding Proteins - metabolism; Humanities and Social Sciences; Hydrogels; Hydrogels - chemistry; Mice; Microenvironments; Microscopy, Electron; Mouse Embryonic Stem Cells - physiology; multidisciplinary; Neuroprotection; Organoids; Organoids - physiology; Organoids - ultrastructure; Photoreceptors; Retina; Retinal Cone Photoreceptor Cells - physiology; Retinal Cone Photoreceptor Cells - ultrastructure; Science; Science (multidisciplinary); Stem cell transplantation; Stem cells; Tissue engineering; Tissue Engineering - methods
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-67012-7

The development of improved methods to culture retinal organoids is relevant for the investigation of mechanisms of retinal development under pathophysiological conditions, for screening of neuroprotective compounds, and for providing a cellular source for clinical transplantation. We report a tissue-engineering approach to accelerate and standardize the production of retinal organoids by culturing mouse embryonic stem cells (mESC) in optimal physico-chemical microenvironments. Arrayed round-bottom milliwells composed of biomimetic hydrogels, combined with an optimized medium formulation, promoted the rapid generation of retina-like tissue from mESC aggregates in a highly efficient and stereotypical manner: ∼93% of the aggregates contained retinal organoid structures. 26 day-old retinal organoids were composed of ∼80% of photoreceptors, of which ∼22% are GNAT2-positive cones, an important and rare sensory cell type that is difficult to study in rodent models. The compartmentalization of retinal organoids into predefined locations on a two-dimensional array not only allowed us to derive almost all aggregates into retinal organoids, but also to reliably capture the dynamics of individual organoids, an advantageous requirement for high-throughput experimentation. Our improved retinal organoid culture system should be useful for applications that require scalability and single-organoid traceability.