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

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 111; no. 23; pp. 8518 - 8523
• Main Authors: Reichman, Sacha, Terray, Angélique, Slembrouck, Amélie, Nanteau, Céline, Orieux, Gaël, Habeler, Walter, Nandrot, Emeline F., Sahel, José-Alain, Monville, Christelle, Goureau, Olivier
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 10.06.2014; National Acad Sciences
• Subjects: Adult; Biological Sciences; Cell Culture Techniques; Cell Differentiation; Cell Line; Cell lines; Cells, Cultured; Cellular biology; Cellular differentiation; Cultured cells; Dermis - cytology; Embryonic stem cells; Epithelial cells; Fibroblasts - cytology; Fibroblasts - metabolism; Gene Expression; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Human health and pathology; Humans; Immunohistochemistry; Induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Life Sciences; Microscopy, Fluorescence; Molecules; Multipotent Stem Cells - cytology; Multipotent Stem Cells - metabolism; Nanog Homeobox Protein; Neural stem cells; Octamer Transcription Factor-3 - genetics; Octamer Transcription Factor-3 - metabolism; Photoreceptor Cells - cytology; Photoreceptor Cells - metabolism; Photoreceptors; Pluripotent stem cells; Retina; Retinal Neurons - cytology; Retinal Neurons - metabolism; Retinal Pigment Epithelium - cytology; Retinal Pigment Epithelium - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Sensory Organs; SOXB1 Transcription Factors - genetics; SOXB1 Transcription Factors - metabolism; Stem cells; Tissues; Transfection; rods; retinal ganglion cells; cones; and O.G. analyzed data; C.N; C.M; A.S; A.T; G.O; W.H; Author contributions: S.R; and O.G. performed research; S.R; and S.R. and O.G. wrote the paper; J.-A.S; and C.M. contributed new reagents/analytic tools; and O.G. designed research; E.F.N
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1324212111

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.