Prof. Marcelo Rivolta
Centre for Stem Cell Biology and Department of Biomedical Sciences, University of Sheffield, UK
Producing and purifying otic progenitors from human pluripotent stem cells: addressing the challenges for their clinical application
The progression of stem cell-derived therapies from lab to bed-side is dependent of adequate, clinical-grade procedures for cell production and quality control. We have developed methods for the efficient production of otic neuroprogenitors from human pluripotent stem cells. However, the protocols currently available produce heterogeneous populations where otic progenitors are invariably mixed with other cell types. Using a reporter human embryonic stem cell line in which GFP is driven by the otic enhancers for SOX2 (NOP1 and NOP2), we have isolated and purified the desired cell population by fluorescence automated cell sorting (FACS). The performance of these cells has been evaluated in vivo and in vitro.
Moreover, an unbiased cell surface antibody screening has been undertaken on otic progenitors derived from hESCs. Screen plates were imaged by automated microscopy thereby maintaining information relating to cell and colony morphologies. This information was used to narrow down potential hits based on known characteristics of otic progenitors. A subset has been selected, potentially enabling the isolation of cells biased to differentiate towards either otic neuronal or inner ear hair cell fates. The refinement and adaption of research grade methods to Good Manufacturing Practice (GMP) standards will enable the translation of these technologies into a clinical application.
Supported by the MRC, the UK Regenerative Medicine Platform and EU-FP7-603029 OtoStem.