Document Detail


Induction of corneal epithelium-like cells from cynomolgus monkey embryonic stem cells and their experimental transplantation to damaged cornea.
MedLine Citation:
PMID:  20164754     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: We previously reported the successful transplantation of corneal epithelium-like cells derived from mouse embryonic stem (ES) cells onto injured mouse cornea. Here, we tested whether nonhuman primate ES cells have ability to differentiate into corneal epithelial cells and whether monkey ES cell-derived corneal epithelium-like cells were applicable for the experimental transplantation to damaged cornea.
METHODS: Monkey ES cells were cultivated on type IV collagen-coated dishes for various days to induce differentiation into corneal epithelium-like cells. The differentiation was evaluated by reverse transcription-polymerase chain reaction and immunostaining. The corneal epithelium-like cells were transplanted to the injured mouse cornea. Reconstitution of the corneal epithelium was evaluated by immunostaining.
RESULTS: The cells cultured on type IV collagen showed cobblestone-like appearance resembling epithelial cells. They expressed messenger RNA of pax6, p63, E-cadherin, CD44, proliferating cell nuclear antigen, keratin 3, and keratin 12. Protein expressions of pax6, keratin 3/12, p63, proliferating cell nuclear antigen, E-cadherin, and CD44 were confirmed by immunostaining. When the corneal epithelium-like cells were transplanted, they adhered to the corneal stroma, leading to formation of multiple cell layers. The grafted cells were stained with anti-human nuclear protein antibody, which cross-reacted with nuclei of monkey cells but not with those of mouse cells. They retained the expressions of keratin 3/12, E-cadherin, and CD44.
CONCLUSIONS: We induced corneal epithelium-like cells from monkey ES cells with moderate efficiency. The cells were successfully transplanted onto the injured mouse cornea. This is the first demonstration that nonhuman primate ES cells were induced to differentiate into corneal epithelium-like cells, which were applicable for transplantation to an animal model of corneal injury.
Authors:
Yuta Kumagai; Manae S Kurokawa; Hiroki Ueno; Maki Kayama; Kazuo Tsubota; Norio Nakatsuji; Yasushi Kondo; Satoki Ueno; Noboru Suzuki
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Cornea     Volume:  29     ISSN:  1536-4798     ISO Abbreviation:  Cornea     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-10-27     Completed Date:  2011-01-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8216186     Medline TA:  Cornea     Country:  United States    
Other Details:
Languages:  eng     Pagination:  432-8     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, St. Marianna University School of Medicine, Kawasaki, Kanagawa, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, CD44 / metabolism
Biological Markers / metabolism
Cadherins / metabolism
Cell Culture Techniques
Cell Differentiation / physiology*
Collagen Type IV / metabolism
Cornea / injuries*
Embryonic Stem Cells / cytology*,  metabolism
Epithelium, Corneal / cytology*,  metabolism
Eye Injuries / metabolism,  pathology,  therapy*
Eye Proteins / metabolism
Female
Homeodomain Proteins / metabolism
Immunohistochemistry
Keratin-12 / metabolism
Keratin-3 / metabolism
Macaca fascicularis
Mice
Mice, Inbred C57BL
Microscopy, Confocal
Paired Box Transcription Factors / metabolism
Proliferating Cell Nuclear Antigen / metabolism
Repressor Proteins / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Stem Cell Transplantation*
Chemical
Reg. No./Substance:
0/Antigens, CD44; 0/Biological Markers; 0/Cadherins; 0/Collagen Type IV; 0/Eye Proteins; 0/Homeodomain Proteins; 0/Keratin-12; 0/Keratin-3; 0/PAX6 protein; 0/Paired Box Transcription Factors; 0/Proliferating Cell Nuclear Antigen; 0/Repressor Proteins

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