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

S Decembrini, S Hoehnel, N Brandenberg, Y Arsenijevic, M P Lutolf

Scientific reports 2020 Jun 24

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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.

Citation

S Decembrini, S Hoehnel, N Brandenberg, Y Arsenijevic, M P Lutolf. Hydrogel-based milliwell arrays for standardized and scalable retinal organoid cultures. Scientific reports. 2020 Jun 24;10(1):10275