Shiro Amano1. 1. Department of Ophthalmology, University of Tokyo School of Medicine, Tokyo, Japan. amanos-tky@umin.ac.jp
Abstract
PURPOSE: To investigate the feasibility of corneal reconstruction utilizing cultured human corneal endothelial cells (HCECs). METHODS: We cultivated HCECs using culture dishes pre-coated with bovine corneal endothelial extracellular matrix. The effect of donor age on HCECs was investigated. We reconstructed corneas using cultured HCECs and human corneal stroma, then examined their functioning. The possibility of porcine corneal stroma as a carrier of cultured HCECs was investigated. RESULTS: The older the donor, the more frequently large senescent cells appeared in the passaged HCECs. The density of HCECs on the reconstructed cornea reached 2500 cells/mm2. The potential difference in the reconstructed and normal corneas was 0.30 mV and 0.40 mV, respectively; this indicates that the pump function of the reconstructed corneas is 75% of that of normal corneas. Porcine corneal stroma expressing little xenosugar antigen alpha-gal epitope induced no superacute rejection but mild cellular rejection when transplanted into corneas of animals possessing natural antibody to alpha-gal epitope. CONCLUSIONS: To reconstruct corneas that are the same as, or superior to, normal corneas, innovation is necessary in the methods used for culturing and seeding HCECs. Porcine corneal stroma is promising as a carrier of HCECs instead of human corneal stroma, the supply of which is limited. The validity of porcine corneal stroma, acellularized to prevent retrovirus infection, should be evaluated.
PURPOSE: To investigate the feasibility of corneal reconstruction utilizing cultured human corneal endothelial cells (HCECs). METHODS: We cultivated HCECs using culture dishes pre-coated with bovine corneal endothelial extracellular matrix. The effect of donor age on HCECs was investigated. We reconstructed corneas using cultured HCECs and humancorneal stroma, then examined their functioning. The possibility of porcine corneal stroma as a carrier of cultured HCECs was investigated. RESULTS: The older the donor, the more frequently large senescent cells appeared in the passaged HCECs. The density of HCECs on the reconstructed cornea reached 2500 cells/mm2. The potential difference in the reconstructed and normal corneas was 0.30 mV and 0.40 mV, respectively; this indicates that the pump function of the reconstructed corneas is 75% of that of normal corneas. Porcine corneal stroma expressing little xenosugar antigen alpha-gal epitope induced no superacute rejection but mild cellular rejection when transplanted into corneas of animals possessing natural antibody to alpha-gal epitope. CONCLUSIONS: To reconstruct corneas that are the same as, or superior to, normal corneas, innovation is necessary in the methods used for culturing and seeding HCECs. Porcine corneal stroma is promising as a carrier of HCECs instead of humancorneal stroma, the supply of which is limited. The validity of porcine corneal stroma, acellularized to prevent retrovirus infection, should be evaluated.
Authors: Giuseppe Orlando; Pedro Baptista; Martin Birchall; Paolo De Coppi; Alan Farney; Nadia K Guimaraes-Souza; Emmanuel Opara; Jeffrey Rogers; Dror Seliktar; Keren Shapira-Schweitzer; Robert J Stratta; Anthony Atala; Kathryn J Wood; Shay Soker Journal: Transpl Int Date: 2010-11-10 Impact factor: 3.782