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- W242089640 abstract "Human embryonic stem cells (HESC) are a promising source of retinal pigment epithelium (RPE) for the treatment of common and incurable forms of blindness such as age-related macular degeneration (AMD). Whilst most HESC lines will produce some pigmented RPE cells when allowed to overgrow and spontaneously differentiate for 30-60 days, the efficiency of this process is highly variable and the critical factors which determine target cell yield remain largely uncharacterised. This will prove problematic in the large-scale production of RPE cells needed for cell therapy. In this project the aim was to identify and minimise sources of variability during differentiation and to develop an efficient and scalable HESC-RPE differentiation protocol. Using a novel imaging platform in combination with quantitative gene expression analysis and immunocytochemistry, the relative differentiation efficiency in two new cell lines, Shef6 and Shef3 was characterised. It was found that the age of the starting HESC population, the cell seeding density and the passaging method used have a strong influence on RPE yields. It was also demonstrated that RPE can be generated from HESC following single cell dissociation and in the absence of feeder cells, thereby significantly simplifying cell culture logistics and reducing variability. In addition it was shown that the lower yielding cell line, Shef3, has a reduced innate propensity for neuroectoderm conversion and that by directing this process with a small molecule, dorsomorphin, efficiency can be significantly improved. Overall this novel protocol increased RPE foci yields per cm2 by 5 fold in Shef6 and 4 fold in Shef3 compared to traditional mouse embryonic fibroblast (MEFs) co-culture systems. Since HESC-derived RPE are now entering clinical trials, it has become increasingly important to optimise manufacturing; this study identifies several critical parameters that could help develop a robust, scalable and cost-effective strategy for HESC-RPE manufacturing." @default.
- W242089640 created "2016-06-24" @default.
- W242089640 creator A5074624696 @default.
- W242089640 date "2014-01-28" @default.
- W242089640 modified "2023-09-28" @default.
- W242089640 title "Bioprocess Design Considerations for the Large Scale Manufacture of Pluripotent Stem Cell Derived Retinal Pigment Epithelium" @default.
- W242089640 hasPublicationYear "2014" @default.
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