1. Differentiation of induced pluripotent stem cells of swine into rod photoreceptors and their integration into the retina.
- Author
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Zhou L, Wang W, Liu Y, Fernandez de Castro J, Ezashi T, Telugu BP, Roberts RM, Kaplan HJ, and Dean DC
- Subjects
- Animals, Antigens, Differentiation metabolism, Cell Culture Techniques, Cells, Cultured, Coculture Techniques, Collagen metabolism, Drug Combinations, Embryoid Bodies cytology, Embryoid Bodies metabolism, Induced Pluripotent Stem Cells metabolism, Induced Pluripotent Stem Cells transplantation, Laminin metabolism, Proteoglycans metabolism, Recoverin metabolism, Retina drug effects, Retinal Rod Photoreceptor Cells metabolism, Retinol-Binding Proteins metabolism, Rhodopsin metabolism, Swine, Tubulin metabolism, Cell Differentiation, Induced Pluripotent Stem Cells cytology, Retina pathology, Retinal Rod Photoreceptor Cells cytology
- Abstract
Absence of a regenerative pathway for damaged retina following injury or disease has led to experiments using stem cell transplantation for retinal repair, and encouraging results have been obtained in rodents. The swine eye is a closer anatomical and physiological match to the human eye, but embryonic stem cells have not been isolated from pig, and photoreceptor differentiation has not been demonstrated with induced pluripotent stem cells (iPSCs) of swine. Here, we subjected iPSCs of swine to a rod photoreceptor differentiation protocol consisting of floating culture as embryoid bodies followed by differentiation in adherent culture. Real-time PCR and immunostaining of differentiated cells demonstrated loss of expression of the pluripotent genes POU5F1, NANOG, and SOX2 and induction of rod photoreceptor genes RCVRN, NRL, RHO, and ROM1. While these differentiated cells displayed neuronal morphology, culturing on a Matrigel substratum triggered a further morphological change resulting in concentration of rhodopsin (RHO) and rod outer segment-specific membrane protein 1 in outer segment-like projections resembling those on primary cultures of rod photoreceptors. The differentiated cells were transplanted into the subretinal space of pigs treated with iodoacetic acid to eliminate rod photoreceptors. Three weeks after transplantation, engrafted RHO+ cells were evident in the outer nuclear layer where photoreceptors normally reside. A portion of these transplanted cells had generated projections resembling outer segments. These results demonstrate that iPSCs of swine can differentiate into photoreceptors in culture, and these cells can integrate into the damaged swine neural retina, thus, laying a foundation for future studies using the pig as a model for retinal stem cell transplantation., (Copyright © 2011 AlphaMed Press.)
- Published
- 2011
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