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Stem Cell Reports. 2018 Jan 9;10(1):300-313. doi: 10.1016/j.stemcr.2017.11.001. Epub 2017 Dec 7.

Accelerated and Improved Differentiation of Retinal Organoids from Pluripotent Stem Cells in Rotating-Wall Vessel Bioreactors.

Author information

1
Neurobiology, Neurodegeneration, and Repair Laboratory (N-NRL), National Eye Institute (NEI), National Institutes of Health, Bldg 6/338, 6 Center Drive, Bethesda, MD 20814, USA.
2
Trans-NIH Shared Resources on Biomedical Engineering and Physical Sciences (BEPS), National Institutes of Biomedical Imaging and Bioengineering (NIBIB), National Institutes of Health, Bldg 13/3N18B, 13 South Drive, Bethesda, MD 20814, USA.
3
Signal Processing and Instrumentation Section, Center for Information Technology (CIT), National Institutes of Health, Bldg 12A/2021, 12 South Drive, Bethesda, MD 20814, USA.
4
Neurobiology, Neurodegeneration, and Repair Laboratory (N-NRL), National Eye Institute (NEI), National Institutes of Health, Bldg 6/338, 6 Center Drive, Bethesda, MD 20814, USA. Electronic address: swaroopa@nei.nih.gov.

Abstract

Pluripotent stem cells can be differentiated into 3D retinal organoids, with major cell types self-patterning into a polarized, laminated architecture. In static cultures, organoid development may be hindered by limitations in diffusion of oxygen and nutrients. Herein, we report a bioprocess using rotating-wall vessel (RWV) bioreactors to culture retinal organoids derived from mouse pluripotent stem cells. Organoids in RWV demonstrate enhanced proliferation, with well-defined morphology and improved differentiation of neurons including ganglion cells and S-cone photoreceptors. Furthermore, RWV organoids at day 25 (D25) reveal similar maturation and transcriptome profile as those at D32 in static culture, closely recapitulating spatiotemporal development of postnatal day 6 mouse retina in vivo. Interestingly, however, retinal organoids do not differentiate further under any in vitro condition tested here, suggesting additional requirements for functional maturation. Our studies demonstrate that bioreactors can accelerate and improve organoid growth and differentiation for modeling retinal disease and evaluation of therapies.

KEYWORDS:

3-D organoid culture; RNA-seq; bioreactor; embryonic stem cell; iPSC; in vitro organogenesis; retina development; retinal disease; transcriptome

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