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Jade Harkin, Kelly Lentsch, Jason S Meyer; Towards a more efficient and reproducible method for retinal organoid specification from human pluripotent stem cells. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1695.
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© ARVO (1962-2015); The Authors (2016-present)
Retinal organoids are three-dimensional aggregates of cells that self-organize into retinal-like tissues. Current protocols utilize human pluripotent stem cells (hPSCs) to generate retinal organoids in vitro, however few protocols have explored the factors that regulate the earliest stages of retinal organoid specification. As such, a more reproducible and efficient method for differentiating early-stage retinal organoids would provide a reliable and reproducible platform for studying human retinogenesis and disease.
In the current study, 3D aggregates of hPSCs were generated using quick reaggregation methods in 96-well U-bottom plates to ensure reproducibility among culture conditions. Various cell densities were tested for the most efficient generation of retinal organoids. Additionally, a role for BMP signaling in retinal organoid differentiation was tested by treatment with either BMP4 or Noggin, and efficiency was assessed at different time points based on morphological analysis and expression of retinal progenitor markers. Ongoing experiments are assessing transcriptional changes that are associated with the specification of a retinal organoid fate.
Results indicate that cell aggregates generated using the quick reaggregation differentiation method are highly reproducible in both their size and shape at early time points compared to traditional methods. Early-stage retinal organoid differentiation using the quick reaggregation method was also significantly improved after BMP4 activation. Ongoing experiments will continue to focus on optimizing current retinal organoid differentiation protocols, as well as identifying the transcriptional alterations that are associated with the development of retinal organoids.
The precise regulation of multiple parameters within this study provides a novel highly reproducible retinal organoid model that can improve our understanding of human retinogenesis as well as retinal degenerative diseases. The efficient generation of reproducible retinal organoids will also accelerate the development of translational approaches for debilitating blinding disorders.
This is a 2021 ARVO Annual Meeting abstract.
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