Open AccessDifferentiation of Induced Pluripotent Stem Cells of Swine into Rod Photoreceptors and Their Integration into the Retina
Author(s) -
Zhou Liang,
Wang Wei,
Liu Yongqing,
de Castro Juan Fernandez,
Ezashi Toshihiko,
Telugu Bhanu Prakash V.L.,
Roberts R. Michael,
Kaplan Henry J.,
Dean Douglas C.
Publication year - 2011
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.637
Subject(s) - biology , induced pluripotent stem cell , microbiology and biotechnology , embryoid body , transplantation , retina , sox2 , embryonic stem cell , stem cell , outer nuclear layer , retinal regeneration , cellular differentiation , homeobox protein nanog , retinal , anatomy , regeneration (biology) , genetics , neuroscience , botany , gene , medicine , surgery
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. STEM CELLS 2011;29:972–980