From confluent human iPS cells to self-forming neural retina and retinal pigmented epithelium

Reichman S, Terray A, Slembrouck A, Nanteau C, Orieux G, Habeler W, Nandrot EF, Sahel JA, Monville C, Goureau O
Source: Proc Natl Acad Sci USA
Publication Date: (2014)
Issue: 111(23): 8518-23
Research Area:
Stem Cells
Cells used in publication:
Fibroblast, dermal(NHDF-Ad), human adult
Species: human
Tissue Origin: dermal
Epithelial, retinal pigment (RPE), human
Species: human
Tissue Origin: eye

adult human fibroblasts 1x10e6/sample, P2, DT-130, co-transfection of 3 plasmids, plated 5x10e4/cm2 on MEF-feeder; after 4 days switch to modified ReproStem media; after 30-40d iPS colonies;


Progress in retinal-cell therapy derived from human pluripotent stem cells currently faces technical challenges that require the development of easy and standardized protocols. Here, we developed a simple retinal differentiation method, based on confluent human induced pluripotent stem cells (hiPSC), bypassing embryoid body formation and the use of exogenous molecules, coating, or Matrigel. In 2 wk, we generated both retinal pigmented epithelial cells and self-forming neural retina (NR)-like structures containing retinal progenitor cells (RPCs). We report sequential differentiation from RPCs to the seven neuroretinal cell types in maturated NR-like structures as floating cultures, thereby revealing the multipotency of RPCs generated from integration-free hiPSCs. Furthermore, Notch pathway inhibition boosted the generation of photoreceptor precursor cells, crucial in establishing cell therapy strategies. This innovative process proposed here provides a readily efficient and scalable approach to produce retinal cells for regenerative medicine and for drug-screening purposes, as well as an in vitro model of human retinal development and disease.