June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Differentiation of human embryonic stem cells to corneal epithelium and endothelium like cells for cornea replacement construction
Author Affiliations & Notes
  • Kunpeng Pang
    Ophthalmology, Qilu Hospital, Shandong University, Jinan, China
  • Kai Zhang
    Ophthalmology, Qilu Hospital, Shandong University, Jinan, China
  • Jing Zhu
    Ophthalmology, Qilu Hospital, Shandong University, Jinan, China
  • Chengqun Ju
    Ophthalmology, Qilu Hospital, Shandong University, Jinan, China
  • Liqun Du
    Ophthalmology, Qilu Hospital, Shandong University, Jinan, China
  • Xinyi Wu
    Ophthalmology, Qilu Hospital, Shandong University, Jinan, China
  • Footnotes
    Commercial Relationships Kunpeng Pang, None; Kai Zhang, None; Jing Zhu, None; Chengqun Ju, None; Liqun Du, None; Xinyi Wu, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 5831. doi:
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      Kunpeng Pang, Kai Zhang, Jing Zhu, Chengqun Ju, Liqun Du, Xinyi Wu; Differentiation of human embryonic stem cells to corneal epithelium and endothelium like cells for cornea replacement construction. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5831.

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

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Abstract

Purpose: A potential application of human embryonic stem cells (hESCs) is the generation of corneal epithelial (CEpi) and endothelial (CEndo) cells to be used for corneal disease treatment. In this study, we designed a new method to induce hESC differentiating to CEpi and CEndo like cells for cornea substitute construction.

Methods: Human embryonic stem cells (hESC) were cultured in a transwell coculture system with differentiated human corneal stromal cells to differentiate into periocular mesenchymal precursor (POMPs). Next, the CEndo-like cells expressed N-cadherin/vimention were derived from POMPs with lens epithelial cell-conditioned medium, and isolated by Fluorescence-activated cell sorting (FACS). To obtain CEpi-like cells, the hESCs were cultured in the limbal stem cells conditioned medium for directional differentiation. The epithelial markers were detected by immunocytochemistry. And then, the induced CEpi and CEndo like cells were cultured on the acellular porcine cornea matrix (APCM) respectively to construct cell sheets. The phenotype of the cell sheet was detected by Immunohistochemistry and electron microscope, and the biological function would be evaluated in vivo.

Results: The CEndo-like cells derived from hESC expressed the corneal endothelium (CE) differentiation marker N-cadherin and transcription factors FoxC1 and Pitx2. The constructed CEndo-like cell sheets had the same pump functions like those of normal corneas. The corneal transparency could be restored gradually after the cell sheets transplanted into the eyes of rabbit CE dysfunction models. And, the induced CEpi -like cells possessed the similar morphologic characteristics and phenotype of normal corneal epithelial cells and showed a strong proliferative capacity in vitro. The CEpi-like cells could form stratified cells sheets on the APCM, and got the ability to reconstruct the damaged ocular surface in cornea alkali burn models.

Conclusions: The method we used was effective to induce hESC differentiating to CEndo and CEpi like cells. And, the induced cells might be used for cornea replacement construction and the treatment of corneal diseases in future.

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