TY  - JOUR
AU  - Völkner, Manuela
AU  - Kurth, Thomas
AU  - Schor, Jana
AU  - Ebner, Lynn J A
AU  - Bardtke, Lara
AU  - Kavak, Cagri
AU  - Hackermüller, Jörg
AU  - Karl, Mike Oliver
TI  - Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture.
JO  - Frontiers in cell and developmental biology
VL  - 9
SN  - 2296-634X
CY  - Lausanne
PB  - Frontiers Media
M1  - DZNE-2021-01265
SP  - 645704
PY  - 2021
N1  - CC BY
AB  - Using retinal organoid systems, organ-like 3D tissues, relies implicitly on their robustness. However, essential key parameters, particularly retinal growth and longer-term culture, are still insufficiently defined. Here, we hypothesize that a previously optimized protocol for high yield of evenly-sized mouse retinal organoids with low variability facilitates assessment of such parameters. We demonstrate that these organoids reliably complete retinogenesis, and can be maintained at least up to 60 days in culture. During this time, the organoids continue to mature on a molecular and (ultra)structural level: They develop photoreceptor outer segments and synapses, transiently maintain its cell composition for about 5-10 days after completing retinogenesis, and subsequently develop pathologic changes - mainly of the inner but also outer retina and reactive gliosis. To test whether this organoid system provides experimental access to the retina during and upon completion of development, we defined and stimulated organoid growth by activating sonic hedgehog signaling, which in patients and mice in vivo with a congenital defect leads to enlarged eyes. Here, a sonic hedgehog signaling activator increased retinal epithelia length in the organoid system when applied during but not after completion of development. This experimentally supports organoid maturation, stability, and experimental reproducibility in this organoid system, and provides a potential enlarged retina pathology model, as well as a protocol for producing larger organoids. Together, our study advances the understanding of retinal growth, maturation, and maintenance, and further optimizes the organoid system for future utilization.
KW  - development (Other)
KW  - gliosis (Other)
KW  - growth (Other)
KW  - maturation (Other)
KW  - mouse (Other)
KW  - organoid (Other)
KW  - retina (Other)
KW  - sonic hedgehog (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:33996806
C2  - pmc:PMC8114082
DO  - DOI:10.3389/fcell.2021.645704
UR  - https://pub.dzne.de/record/157808
ER  -