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ICE Krakow

ICE Krakow

ul. Marii Konopnickiej 17 30-302 Kraków


Jongen, Vincent (Masaryk University )


Advances in organoid research has led to the development of retinal organoids capable of forming optic cups as well as giving rise to all major cell types of the neuroretina, most notably mature photoreceptors. These retinal organoids can be used for studying retinal development or to serve as a source of photoreceptors for transplantation therapy. However, several challenges remain, such as heterogeneity between organoids, poor photoreceptor maturation and the degradation of retinal ganglion cells in long term cultures. Microfluidic platforms have the potential to help overcome some of these challenges by allowing for automation and precise control of the microenvironment. For this work we designed a one-stop microfluidic device capable of retinal organoid formation and maturation to decrease labor intensity while improving organoid oxygen and nutrient supply.

Microfluidic devices were fabricated from PDMS using a mold created with photo- and stereolithography. Retinal organoids were differentiated from human pluripotent stem cells inside a microfluidic device under continuous medium perfusion. The organoids were compared to a static control, grown in standard culture conditions, based on the analysis of morphology, cell viability, and cell differentiation.

Optimization of the shape and size of the microwells allowed for efficient formation of homogenous embryoid bodies within 24 hours after seeding. Subsequent cultivation under continuous medium perfusion led to the development of retinal organoids, as observed using live imaging. Retinal organoids cultivated within the chip developed at a timescale representative of the in vivo situation as it was evaluated by the expression of eye field transcription markers and retinal progenitor cell markers.

The proposed platform allows for standardized formation and long-term maturation of retinal organoids in a low shear-stress environment while decreasing labor intensity. An array of microwells allowed for quick aggregation and organoid formation. The device was capable of automated continuous medium perfusion to allow for retinal organoid cultivation and maturation without the need for transferring the organoids to a secondary culture vessel.

This work was supported by the European Regional Development Fund (project INBIO CZ.02.1.01/0.0/0.0/16_026/0008451), Internal Grant Agency of Masaryk University (project MUNI/IGA/1297/2021 and MUNI/A/1398/2021), Brno city municipality (Brno Ph.D. Talent Scholarship), Ministry of Health of the Czech Republic (NU21-08-00561)


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