Fidelity in pluripotent stem cell differentiation protocols is necessary for the therapeutic and commercial use of cells derived from embryonic and induced pluripotent stem cells. Recent advances in stem cell technology, especially the widespread availability of a range of chemically defined media, substrates and differentiation components, now allow the design and implementation of fully defined derivation and differentiation protocols intended for replication across multiple research and manufacturing locations. In this report we present an application of these criteria to the generation of retinal pigmented epithelium from iPSCs derived from the conjunctiva of donors with and without age related macular degeneration. Primary conjunctival cells from human donors aged 70-85 years were reprogrammed to derive multiple iPSC lines that were differentiated into functional RPE using a rapid and defined differentiation protocol. The combination of defined iPSC derivation and culture with a defined RPE differentiation protocol, reproducibly generated functional RPE from each donor without requiring protocol adjustments for each individual. This successful validation of a standardized, iPSC derivation and RPE differentiation process demonstrates a practical approach for applications requiring the cost-effective generation of RPE from multiple individuals such as drug testing, population studies or for therapies requiring patient-specific RPE derivations. In addition, conjunctival cells are identified as a practical source of somatic cells for deriving iPSCs from elderly individuals.
Bibliographical noteFunding Information:
This work was supported by funds from the Elaine and Robert Larson Endowed Vision Research Chair (to DAF) and an anonymous donor for Macular Degeneration Research. This work was supported in part by a grant from Regenerative Medicine Minnesota (MRM 2015 5337 to DAF and JRD), an unrestricted grant from Research to Prevent Blindness to the Department of Ophthalmology and Visual Neurosciences and the Core Grant for Vision Research from the National Eye Institute (P30-EY11374). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
© 2017 Geng et al.