June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Corneal endothelial cells derived from monkey iPS cells: a short term evaluation
Author Affiliations & Notes
  • Shin Hatou
    Department of Ophthalmology, Keio Univ School of Medicine, Shinjuku-ku, Japan
  • Satoru Yoshida
    Department of Ophthalmology, Keio Univ School of Medicine, Shinjuku-ku, Japan
  • Kazunari Higa
    Department of Ophthalmology, Tokyo Dental College Ichikawa General Hospital, Ichikawa, Japan
  • Hideyuki Miyashita
    Department of Ophthalmology, Keio Univ School of Medicine, Shinjuku-ku, Japan
  • Emi Inagaki
    Department of Ophthalmology, Keio Univ School of Medicine, Shinjuku-ku, Japan
  • Erika Kimura
    Department of Ophthalmology, Osaka University, Osaka, Japan
  • Ryuhei Hayashi
    Department of Ophthalmology, Osaka University, Osaka, Japan
  • Kazuo Tsubota
    Department of Ophthalmology, Keio Univ School of Medicine, Shinjuku-ku, Japan
  • Kohji Nishida
    Department of Ophthalmology, Osaka University, Osaka, Japan
  • Shigeto Shimmura
    Department of Ophthalmology, Keio Univ School of Medicine, Shinjuku-ku, Japan
  • Footnotes
    Commercial Relationships Shin Hatou, None; Satoru Yoshida, None; Kazunari Higa, None; Hideyuki Miyashita, None; Emi Inagaki, None; Erika Kimura, None; Ryuhei Hayashi, None; Kazuo Tsubota, AcuFocus, Inc (C), Allergan (F), Bausch Lomb Surgical (C), Functional visual acuity meter (P), JiNS (P), Kissei (F), Kowa (F), Santen, Inc. (F), Otsuka (F), Pfizer (C), Thea (C), Echo Denki (P), Nidek (F), Ophtecs (F), Wakasa Seikatsu (F), CEPT Company (P); Kohji Nishida, Alcon (C), Alcon (F), HOYA (F), Senju (F), Pfizer (F), Santen (F), Osaka University (P); Shigeto Shimmura, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1015. doi:
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      Shin Hatou, Satoru Yoshida, Kazunari Higa, Hideyuki Miyashita, Emi Inagaki, Erika Kimura, Ryuhei Hayashi, Kazuo Tsubota, Kohji Nishida, Shigeto Shimmura; Corneal endothelial cells derived from monkey iPS cells: a short term evaluation. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1015.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: To evaluate the short term function of tissue-engineered corneal endothelial cells (TECE cells) derived from monkey induced pluripotent stem cells (iPS cells).

Methods: Cynomolgus monkey iPS cells were cultured in KSR medium for 1 week and subsequently in N2 medium for 1 week, supplemented with TGF-beta inhibitor and BMP inhibitor. The iPS-derived neural crest cells were isolated as CD271 positive fraction by cell sorter. Next these cells were proliferated with EGF and FGF2, and subsequently medium was changed to an “endothelium-deriving medium” including GSK-3beta inhibitor, retinoic acid and ROCK inhibitor. These cells were dispersed on collagen sheet and TECE cell sheets were obtained. The pump function attributable to Na,K-ATPase activity of TECE cell sheets was measured with an Ussing chamber, and compared with that of human corneal endothelial cell line (B4G12 cells). In vivo function of TECE was measured as central corneal thickness of rabbit eyes transplanted with TECE cell sheets for 8 days after surgery and compared with control eyes deprived of endothelium.

Results: Hexagonal mosaic pattern monolayer TECE cells were obtained. Pump function of TECE was 2.34±0.46 mV, whereas that of B4G12 cells was 1.07±0.20 mV. The corneal thickness of TECE transplanted rabbit eyes (589.25±164.8μm) maintained significant lower corneal thickness than control eyes (1105.8±165.9μm) throughout the post-operative period.

Conclusions: In vitro and short term in vivo function of monkey iPS-derived TECE were observed. Further long-term in vivo evaluation of TECE transplantation to monkey eyes is needed.

Keywords: 481 cornea: endothelium • 721 stem cells • 606 NaK ATPase  
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