Induction of corneal epithelium-like cells from cynomolgus monkey embryonic stem cells and their experimental transplantation to damaged cornea

Yuta Kumagai, Manae S. Kurokawa, Hiroki Ueno, Maki Kayama, Kazuo Tsubota, Norio Nakatsuji, Yasushi Kondo, Satoki Ueno, Noboru Suzuki

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

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 crossreacted 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.

Original languageEnglish
Pages (from-to)432-438
Number of pages7
JournalCornea
Volume29
Issue number4
DOIs
Publication statusPublished - 2010 Apr

Fingerprint

Corneal Epithelium
Macaca fascicularis
Embryonic Stem Cells
Cornea
Transplantation
Keratin-12
Keratin-3
Haplorhini
Cadherins
Collagen Type IV
Proliferating Cell Nuclear Antigen
Primates
Epithelial Cells
Corneal Stroma
Nuclear Proteins
Cell Nucleus
Reverse Transcription
Cultured Cells
Animal Models

Keywords

  • collagen type IV
  • cornea
  • epithelial cells
  • keratin 12
  • monkey embryonic stem cell

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Induction of corneal epithelium-like cells from cynomolgus monkey embryonic stem cells and their experimental transplantation to damaged cornea. / Kumagai, Yuta; Kurokawa, Manae S.; Ueno, Hiroki; Kayama, Maki; Tsubota, Kazuo; Nakatsuji, Norio; Kondo, Yasushi; Ueno, Satoki; Suzuki, Noboru.

In: Cornea, Vol. 29, No. 4, 04.2010, p. 432-438.

Research output: Contribution to journalArticle

Kumagai, Yuta ; Kurokawa, Manae S. ; Ueno, Hiroki ; Kayama, Maki ; Tsubota, Kazuo ; Nakatsuji, Norio ; Kondo, Yasushi ; Ueno, Satoki ; Suzuki, Noboru. / Induction of corneal epithelium-like cells from cynomolgus monkey embryonic stem cells and their experimental transplantation to damaged cornea. In: Cornea. 2010 ; Vol. 29, No. 4. pp. 432-438.
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AU - Kayama, Maki

AU - Tsubota, Kazuo

AU - Nakatsuji, Norio

AU - Kondo, Yasushi

AU - Ueno, Satoki

AU - Suzuki, Noboru

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N2 - 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 crossreacted 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.

AB - 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 crossreacted 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.

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KW - epithelial cells

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