Effects of extracellular matrix and neighboring cells on induction of human embryonic stem cells into retinal or retinal pigment epithelial progenitors.

To determine the effects of extracellular matrix and neighboring cells on the differentiation of human embryonic stem cells (hESC) into progenitors of retinal cells and/or retinal pigment epithelium (RPE). HESC were cultured on mouse PA6 stromal cells for approximately 2weeks to obtain neural progenitors. To induce photoreceptor marker expression, the neural progenitors were cultured on a confluent monolayer of ARPE19 or on laminin-coated dishes. To induce RPE markers, the neural progenitors were seeded onto human Bruch's membrane or Matrigel. Cells were examined morphologically and stained with different RPE or neural progenitor markers. Microarray techniques were used to compare the gene expression profiles of hESC cultured on mouse fibroblasts or neural progenitors on PA6 cells to the transcriptome of the adult neural retina and RPE. HESC cultured on PA6 cells expressed neural progenitor markers beta-tubulin III, PAX6, neural filament, GFAP and vimentin. Culturing these neural progenitors on confluent ARPE19 monolayer induced expression of the photoreceptor progenitor cell marker CRX; culturing neural progenitors on laminin substrates induced a neuronal phenotype with neurite formation. Neural progenitors expressed the RPE marker ZO-1 after culturing on Matrigel-coated dishes and the RPE marker Bestrophin after culturing on human Bruch's membrane explants. Hierarchical clustering analysis of samples suggested that when cultured on PA6 stromal cells hESC exhibited genetic characteristics towards differentiating into neural retina. Microarray analysis showed that after culturing on PA6 cells, stem cells expressed 117 new genes; among these there were 22 genes present in neural retina or RPE cells. The functions of these genes were highly related to cell proliferation, nervous system development and cell adhesion. HESC can be induced to differentiate into neural progenitors after culturing on PA6 cells. These neural progenitors can express RPE markers when cultured on Bruch's membrane or Matrigel, or photoreceptor markers when cultured on confluent ARPE19 or laminin. Additional studies are required to assess the function of hESC induced to express retinal or RPE markers prior to successful intraocular transplantation into animal models of retinal degeneration.

[1]  P. Layer,et al.  Inductive effects of the retinal pigmented epithelium (RPE) on histogenesis of the avian retina as revealed by retinospheroid technology. , 1998, Seminars in cell & developmental biology.

[2]  Kenji Mizuseki,et al.  Generation of dopaminergic neurons and pigmented epithelia from primate ES cells by stromal cell-derived inducing activity , 2002, Proceedings of the National Academy of Sciences of the United States of America.

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

[4]  M. Zarbin,et al.  Progressive presumed choriocapillaris atrophy after surgery for age-related macular degeneration. , 1998, Retina.

[5]  M. Schwartz,et al.  The regulation of growth and intracellular signaling by integrins. , 1996, Endocrine reviews.

[6]  P. Henkind,et al.  The relationship between retinal pigment epithelium and the choriocapillaris. , 1983, Transactions of the ophthalmological societies of the United Kingdom.

[7]  D. Sheppard,et al.  Essential role for MFG-E8 as ligand for αvβ5 integrin in diurnal retinal phagocytosis , 2007, Proceedings of the National Academy of Sciences.

[8]  P. Gouras,et al.  Changes in the choroidal circulation of rabbit following RPE removal , 2003, Graefe's Archive for Clinical and Experimental Ophthalmology.

[9]  Y. Sasai,et al.  Optimal conditions for in vivo induction of dopaminergic neurons from embryonic stem cells through stromal cell‐derived inducing activity , 2002, Journal of neuroscience research.

[10]  T. Léveillard,et al.  [Retinal repair by transplantation of photoreceptor precursors]. , 2007, Medecine sciences : M/S.

[11]  H. Kaplan,et al.  Pathogenesis of AMD. , 1995, Ophthalmology.

[12]  J. Frade,et al.  Developmentally regulated vitronectin influences cell differentiation, neuron survival and process outgrowth in the developing chicken retina , 1995, Neuroscience.

[13]  Y. Sasai,et al.  Neural conversion of ES cells by an inductive activity on human amniotic membrane matrix. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Henkind,et al.  Choriocapillaris atrophy after experimental destruction of the retinal pigment epithelium in the rat. A study in thin sections and vascular casts. , 1986, Acta anatomica.

[15]  G. Martin,et al.  Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[16]  H. Kaplan,et al.  Débridement of the pig retinal pigment epithelium in vivo. , 1995, Archives of ophthalmology.

[17]  Y. Sasai,et al.  Generation of graftable dopaminergic neuron progenitors from mouse ES cells by a combination of coculture and neurosphere methods , 2006, Journal of neuroscience research.

[18]  Y. Sasai,et al.  In vitro and in vivo characterization of pigment epithelial cells differentiated from primate embryonic stem cells. , 2004, Investigative ophthalmology & visual science.

[19]  H. Kaplan,et al.  Postoperative abnormalities of the choriocapillaris in exudative age-related macular degeneration. , 1996, The British journal of ophthalmology.

[20]  H. Kaplan,et al.  Choriocapillaris atrophy after submacular surgery in presumed ocular histoplasmosis syndrome. , 1995, Archives of ophthalmology.

[21]  S. Saule,et al.  Microphthalmia Transcription Factor Induces Both Retinal Pigmented Epithelium and Neural Crest Melanocytes from Neuroretina Cells* , 2004, Journal of Biological Chemistry.

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

[23]  K. Mizuseki,et al.  Generation of Rx+/Pax6+ neural retinal precursors from embryonic stem cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Jian-xing Ma,et al.  Isorhodopsin rather than rhodopsin mediates rod function in RPE65 knock-out mice , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. Zarbin,et al.  Decreased choriocapillaris perfusion following surgical excision of choroidal neovascular membranes in age-related macular degeneration , 1997, The British journal of ophthalmology.

[26]  H. Kaplan,et al.  Perfusion of the subfoveal choriocapillaris affects visual recovery after submacular surgery in presumed ocular histoplasmosis syndrome. , 1997, American journal of ophthalmology.

[27]  C. Cepko.  The roles of intrinsic and extrinsic cues and bHLH genes in the determination of retinal cell fates , 1999, Current Opinion in Neurobiology.

[28]  D. Sheppard,et al.  Essential role for MFG-E8 as ligand for alphavbeta5 integrin in diurnal retinal phagocytosis. , 2007, Proceedings of the National Academy of Sciences of the United States of America.

[29]  H. Kaplan,et al.  Reengineering of aged Bruch's membrane to enhance retinal pigment epithelium repopulation. , 2004, Investigative ophthalmology & visual science.

[30]  T. Kunisada,et al.  Induction of melanocytes from embryonic stem cells and their therapeutic potential. , 2006, Pigment cell research.

[31]  M. Silverman,et al.  Retinal pigment epithelial repopulation in monkeys after submacular surgery. , 1995, Archives of ophthalmology.

[32]  K. Preissner,et al.  Vitronectin gene expression in the adult human retina. , 1999, Investigative ophthalmology & visual science.

[33]  M. Li,et al.  Early specification of dopaminergic phenotype during ES cell differentiation , 2007, BMC Developmental Biology.

[34]  M. Bissell,et al.  The Influence of Extracellular Matrix on Gene Expression: Is Structure the Message? , 1987, Journal of Cell Science.

[35]  V. Reppucci,et al.  RPE destruction causes choriocapillary atrophy. , 1984, Investigative ophthalmology & visual science.

[36]  G. Korte.  Choriocapillaris regeneration in the rabbit. Ultrastructure of new endothelial tube formation. , 1989, Investigative ophthalmology & visual science.

[37]  D. Krause,et al.  Regulation of hematopoietic stem cell fate , 2002, Oncogene.

[38]  H. Okazaki,et al.  High invasiveness associated with augmentation of motility in a fos-transferred highly metastatic rat 3Y1 cell line. , 1989, Cancer research.

[39]  L. Trusolino,et al.  Interactions between growth factor receptors and adhesion molecules: breaking the rules. , 2003, Current opinion in cell biology.

[40]  E. Rodriguez-Boulan,et al.  Phagocytosis of rod outer segments by retinal pigment epithelial cells requires αvβ5 integrin for binding but not for internalization , 1997 .

[41]  R. Langer,et al.  Creating an Immune‐Privileged Site Using Retinal Progenitor Cells and Biodegradable Polymers , 2007, Stem cells.

[42]  Y. Sasai.  Directed differentiation of neural and sensory tissues from embryonic stem cells in vitro. , 2005, Ernst Schering Research Foundation workshop.

[43]  池田 華子.  Generation of Rx[+]/Pax6[+] neural retinal precursors from embryonic stem cells , 2008 .

[44]  H. Kaplan,et al.  Use of iris pigment epithelium to replace retinal pigment epithelium in age-related macular degeneration: a gene expression analysis. , 2006, Archives of ophthalmology.

[45]  K. Mizuseki,et al.  Induction of Midbrain Dopaminergic Neurons from ES Cells by Stromal Cell–Derived Inducing Activity , 2000, Neuron.

[46]  G. Chader,et al.  Matrigel promotes retinoblastoma cell growth in vitro and in vivo , 1992, International journal of cancer.

[47]  L. D. Del Priore,et al.  Bruch Membrane Aging Alters the Gene Expression Profile of Human Retinal Pigment Epithelium , 2006, Current eye research.

[48]  C. Cepko,et al.  Notch 1 inhibits photoreceptor production in the developing mammalian retina , 2006, Development.

[49]  Alon Spira,et al.  High-Resolution Electrophysiological Assessment of Human Embryonic Stem Cell-Derived Cardiomyocytes: A Novel In Vitro Model for the Study of Conduction , 2002, Circulation research.

[50]  H. Kaplan,et al.  Retinal pigment epithelium rescues vascular endothelium from retinoic acid-induced apoptosis. , 2006, Investigative ophthalmology & visual science.

[51]  I. Jackson,et al.  The retinal pigmented epithelium is required for development and maintenance of the mouse neural retina , 1995, Current Biology.

[52]  D. Newsome,et al.  Vitronectin is responsible for serum-stimulated uptake of rod outer segments by cultured retinal pigment epithelial cells. , 1997, Investigative ophthalmology & visual science.

[53]  A. Iannaccone,et al.  Pigment Epithelium-Derived Factor Supports Normal Development of Photoreceptor Neurons and Opsin Expression after Retinal Pigment Epithelium Removal , 2000, The Journal of Neuroscience.

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

[55]  E. Rodriguez-Boulan,et al.  Phagocytosis of rod outer segments by retinal pigment epithelial cells requires alpha(v)beta5 integrin for binding but not for internalization. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[56]  M. Zarbin,et al.  Clinicopathological correlation of an excised choroidal neovascular membrane in pseudotumour cerebri , 1997, The British journal of ophthalmology.

[57]  M. Zarbin,et al.  Clinicopathologic correlation of localized retinal pigment epithelium debridement. , 1997, Investigative ophthalmology & visual science.

[58]  H. Kaplan,et al.  Retinal pigment epithelial debridement as a model for the pathogenesis and treatment of macular degeneration. , 1996, American journal of ophthalmology.