Use of bioreactors for culturing human retinal organoids improves photoreceptor yields
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RESEARCH
Open Access
Use of bioreactors for culturing human retinal organoids improves photoreceptor yields Patrick Ovando-Roche1, Emma L. West1, Matthew J. Branch1, Robert D. Sampson1, Milan Fernando1, Peter Munro1, Anastasios Georgiadis1, Matteo Rizzi1, Magdalena Kloc1, Arifa Naeem1, Joana Ribeiro1, Alexander J. Smith1, Anai Gonzalez-Cordero1 and Robin R. Ali1,2*
Abstract Background: The use of human pluripotent stem cell-derived retinal cells for cell therapy strategies and disease modelling relies on the ability to obtain healthy and organised retinal tissue in sufficient quantities. Generating such tissue is a lengthy process, often taking over 6 months of cell culture, and current approaches do not always generate large quantities of the major retinal cell types required. Methods: We adapted our previously described differentiation protocol to investigate the use of stirred-tank bioreactors. We used immunohistochemistry, flow cytometry and electron microscopy to characterise retinal organoids grown in standard and bioreactor culture conditions. Results: Our analysis revealed that the use of bioreactors results in improved laminar stratification as well as an increase in the yield of photoreceptor cells bearing cilia and nascent outer-segment-like structures. Conclusions: Bioreactors represent a promising platform for scaling up the manufacture of retinal cells for use in disease modelling, drug screening and cell transplantation studies. Keywords: Pluripotent stem cells, Retinal organoids, Bioreactors, Photoreceptors
Background Visual impairment caused by inherited retinal degenerations as well as more complex heterogeneous retinal diseases such as age-related macular degeneration (AMD) and glaucoma are mainly due to the dysfunction or loss of key retinal cells such as retinal pigment epithelium (RPE) cells, photoreceptors or retinal ganglion cells (RGCs). Whilst animal models have provided many important insights into retinal disorders, they often do not recapitulate key aspects of human pathophysiology and, as a result, the molecular mechanisms underlying retinal dystrophies remain poorly understood. This has hampered the development of target-based drug screening strategies employed by the pharmaceutical industry. * Correspondence: [email protected] 1 Department of Genetics, UCL Institute of Ophthalmology, 11–43 Bath Street, London EC1V 9EL, UK 2 NIHR Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, City Road, London EC1V 2PD, UK
The discovery of somatic cell reprogramming has enabled the generation of induced pluripotent stem cells (iPSCs) from adult tissue and allowed the use of patient-derived cells for in-vitro modelling of retinal disorders and cell replacement therapy (for reviews see [1–3]). Furthermore, CRISPR/Cas9 gene editing technology can be harnessed to repair the disease-causing mutation in patient-derived human iPSCs (hiPSCs) to generate unaffected and affected isogenic cell line pairs that will be critical for precise disease
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