Optic Vesicle‐like Structures Derived from Human Pluripotent Stem Cells Facilitate a Customized Approach to Retinal Disease Treatment

Differentiation methods for human induced pluripotent stem cells (hiPSCs) typically yield progeny from multiple tissue lineages, limiting their use for drug testing and autologous cell transplantation. In particular, early retina and forebrain derivatives often intermingle in pluripotent stem cell cultures, owing to their shared ancestry and tightly coupled development. Here, we demonstrate that three‐dimensional populations of retinal progenitor cells (RPCs) can be isolated from early forebrain populations in both human embryonic stem cell and hiPSC cultures, providing a valuable tool for developmental, functional, and translational studies. Using our established protocol, we identified a transient population of optic vesicle (OV)‐like structures that arose during a time period appropriate for normal human retinogenesis. These structures were independently cultured and analyzed to confirm their multipotent RPC status and capacity to produce physiologically responsive retinal cell types, including photoreceptors and retinal pigment epithelium (RPE). We then applied this method to hiPSCs derived from a patient with gyrate atrophy, a retinal degenerative disease affecting the RPE. RPE generated from these hiPSCs exhibited a disease‐specific functional defect that could be corrected either by pharmacological means or following targeted gene repair. The production of OV‐like populations from human pluripotent stem cells should facilitate the study of human retinal development and disease and advance the use of hiPSCs in personalized medicine. STEM CELLS 2011;29:1206‐1218

[1]  G. Addison.  The Metabolic and Molecular Bases of Inherited Disease CD-ROM (Version 1.0) Edited by C.R. Scriver, A.L. Beaudet, W.S. Sly and D. Valle , 1997, Journal of Inherited Metabolic Disease.

[2]  P. Gruss,et al.  Generating neuronal diversity in the retina: one for nearly all , 2002, Trends in Neurosciences.

[3]  Shinya Yamanaka,et al.  Induced pluripotent stem cells and reprogramming: seeing the science through the hype , 2009, Nature Reviews Genetics.

[4]  Z. Kozmík,et al.  Spatial and temporal regulation of Wnt/beta-catenin signaling is essential for development of the retinal pigment epithelium. , 2009, Developmental biology.

[5]  H. Ichinose,et al.  Suppression by an h current of spontaneous Na+ action potentials in human cone and rod photoreceptors. , 2005, Investigative ophthalmology & visual science.

[6]  James A. Thomson,et al.  Induced pluripotent stem cells from a spinal muscular atrophy patient , 2009, Nature.

[7]  R. Lang,et al.  Early eye development in vertebrates. , 2001, Annual review of cell and developmental biology.

[8]  Tamar Dvash,et al.  Molecular signature of primary retinal pigment epithelium and stem-cell-derived RPE cells. , 2010, Human molecular genetics.

[9]  G. Shepherd,et al.  Retinal ganglion cells express a cGMP-gated cation conductance activatable by nitric oxide donors , 1994, Neuron.

[10]  D. Valle,et al.  A mouse model of gyrate atrophy of the choroid and retina. Early retinal pigment epithelium damage and progressive retinal degeneration. , 1996, The Journal of clinical investigation.

[11]  P. Gruss,et al.  Current views on eye development , 1997, Trends in Neurosciences.

[12]  J. I. Korenbrot,et al.  Permeation and interaction of monovalent cations with the cGMP-gated channel of cone photoreceptors , 1992, The Journal of general physiology.

[13]  C. Cepko,et al.  Transdifferentiation of the retina into pigmented cells in ocular retardation mice defines a new function of the homeodomain gene Chx10 , 2004, Development.

[14]  B. Finlay.  The developing and evolving retina: Using time to organize form , 2008, Brain Research.

[15]  Jason Hipp,et al.  Derivation and comparative assessment of retinal pigment epithelium from human embryonic stem cells using transcriptomics. , 2004, Cloning and stem cells.

[16]  D. Clegg,et al.  Derivation of Functional Retinal Pigmented Epithelium from Induced Pluripotent Stem Cells , 2009, Stem cells.

[17]  C. Obie,et al.  Pyridoxine-responsive gyrate atrophy of the choroid and retina: clinical and biochemical correlates of the mutation A226V. , 1995, American journal of human genetics.

[18]  M. Berridge,et al.  Elementary and global aspects of calcium signalling. , 1997, The Journal of experimental biology.

[19]  S. Ito,et al.  Prevention of ornithine cytotoxicity by proline in human retinal pigment epithelial cells. , 1998, Investigative ophthalmology & visual science.

[20]  Fumitaka Osakada,et al.  Modeling Retinal Degeneration Using Patient-Specific Induced Pluripotent Stem Cells , 2011, PloS one.

[21]  T. Adachi,et al.  Self-organizing optic-cup morphogenesis in three-dimensional culture , 2011, Nature.

[22]  Su-Chun Zhang,et al.  Modeling early retinal development with human embryonic and induced pluripotent stem cells , 2009, Proceedings of the National Academy of Sciences.

[23]  D. Valle,et al.  Correction of ornithine accumulation prevents retinal degeneration in a mouse model of gyrate atrophy of the choroid and retina. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[24]  K. Hochedlinger,et al.  Induced pluripotency: history, mechanisms, and applications. , 2010, Genes & development.

[25]  Y. Barishak.  Embryology of the eye and its adnexae. , 1992, Developments in ophthalmology.

[26]  S. Cole,et al.  Sequences Human Induced Pluripotent Stem Cells Free of Vector and Transgene , 2012 .

[27]  Nagahisa Yoshimura,et al.  Toward the generation of rod and cone photoreceptors from mouse, monkey and human embryonic stem cells , 2008, Nature Biotechnology.

[28]  A. Trounson,et al.  Human embryonic stem cells: prospects for development , 2004, Development.

[29]  C. Barnstable,et al.  Molecular aspects of vertebrate retinal development , 2002, Molecular Neurobiology.

[30]  Krishanu Saha,et al.  Pluripotency and Cellular Reprogramming: Facts, Hypotheses, Unresolved Issues , 2010, Cell.

[31]  K. Yau,et al.  Phototransduction Motifs and Variations , 2009, Cell.

[32]  Pei-Rong Wang,et al.  Generation, Purification and Transplantation of Photoreceptors Derived from Human Induced Pluripotent Stem Cells , 2010, PloS one.

[33]  Sheldon S Miller,et al.  Extracellular ATP Activates Calcium Signaling, Ion, and Fluid Transport in Retinal Pigment Epithelium , 1997, The Journal of Neuroscience.

[34]  S. Picaud,et al.  GABAA and GABAC receptors in adult porcine cones: evidence from a photoreceptor‐glia co‐culture model , 1998, The Journal of physiology.

[35]  T. Ben-Hur,et al.  Retinal Incorporation and Differentiation of Neural Precursors Derived from Human Embryonic Stem Cells , 2006, Stem cells.

[36]  T. Reh,et al.  Microarray characterization of human embryonic stem cell--derived retinal cultures. , 2011, Investigative ophthalmology & visual science.

[37]  James A Thomson,et al.  Neural differentiation of human induced pluripotent stem cells follows developmental principles but with variable potency , 2010, Proceedings of the National Academy of Sciences.

[38]  J. Sahel,et al.  Glycine receptors in a population of adult mammalian cones , 2006, The Journal of physiology.

[39]  R. Mcinnes,et al.  Chx10 repression of Mitf is required for the maintenance of mammalian neuroretinal identity , 2004, Development.

[40]  D. Baylor,et al.  Cyclic GMP-activated conductance of retinal photoreceptor cells. , 1989, Annual review of neuroscience.

[41]  A. Menini.  Cyclic nucleotide-gated channels in visual and olfactory transduction. , 1995, Biophysical chemistry.

[42]  Shulan Tian,et al.  Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells , 2007, Science.

[43]  R. Stewart,et al.  Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences , 2009, Science.

[44]  Jason S. Meyer,et al.  Directed differentiation of human induced pluripotent stem cells: a retina perspective. , 2010, Regenerative medicine.

[45]  C. Svendsen,et al.  A novel serum-free method for culturing human prenatal retinal pigment epithelial cells. , 2008, Investigative ophthalmology & visual science.

[46]  K. Yau,et al.  Phototransduction mechanism in retinal rods and cones. The Friedenwald Lecture. , 1994, Investigative ophthalmology & visual science.

[47]  J. Sahel,et al.  GABAC Receptors Are Localized with Microtubule-Associated Protein 1B in Mammalian Cone Photoreceptors , 2000, The Journal of Neuroscience.

[48]  C. Ware,et al.  Efficient generation of retinal progenitor cells from human embryonic stem cells , 2006, Proceedings of the National Academy of Sciences.

[49]  Brian E. McIntosh,et al.  Genetic correction and analysis of induced pluripotent stem cells from a patient with gyrate atrophy , 2011, Proceedings of the National Academy of Sciences.

[50]  R. Kageyama,et al.  Retinal cell fate determination and bHLH factors. , 2004, Seminars in cell & developmental biology.

[51]  G. Keller,et al.  Embryonic stem cell differentiation: emergence of a new era in biology and medicine. , 2005, Genes & development.

[52]  T. Reh,et al.  Extraocular mesenchyme patterns the optic vesicle during early eye development in the embryonic chick. , 2000, Development.

[53]  J. Sahel,et al.  Purification of mammalian cone photoreceptors by lectin panning and the enhancement of their survival in glia-conditioned medium. , 2005, Investigative ophthalmology & visual science.

[54]  Gideon Rechavi,et al.  Directed differentiation of human embryonic stem cells into functional retinal pigment epithelium cells. , 2009, Cell stem cell.

[55]  George Q. Daley,et al.  Disease-Specific Induced Pluripotent Stem Cells , 2008, Cell.

[56]  M. Nguyen,et al.  Signaling and transcriptional regulation in early mammalian eye development: a link between FGF and MITF. , 2000, Development.

[57]  Wenfang Liu,et al.  Alternative promoter use in eye development: the complex role and regulation of the transcription factor MITF , 2008, Development.