Corneal Reconstruction by tissue-engineering

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The older the donor was, the more frequently large senescent cells appeared in the passaged HCEC. HCEC was cultured using adult human serum instead of bovine serum to get rid of bovine material that can be infected with prion. Primary and passage culture of HCEC was possible using adult human serum. We reconstructed the cornea using cultured HCEC and human corneal stroma. The corneal stroma, on which cell suspension of HCEC was poured, was mildly centrifuged to enhance the HCEC attachment to the stroma. The cell density of HCEC on the reconstructed cornea reached to 2500 cells/mm^2. The pump function of the reconstructed cornea was measured with Ussing chamber The potential difference in the reconstructed cornea and normal cornea was 0.30 mV and 0.40 mV, respectively ; indicating the pump function of the reconstructed cornea is 75% of that of the normal cornea. The reconstructed cornea was transplanted to rabbit eye and stayed transparent during postoperative 6 months. Fluorescein labeled cultured HCEC remained on the graft 1 month after the transplantation, indicating transplanted HCEC contributed to the transparency of the graft. The artificial stroma made of alkaline-treated collagen could not be sutured but showed good transparency, biocompatibility, and cell-attachability. Porcine corneal stroma, expressing little xeno-sugar antigen α-gal epitope, induced no super acute rejection but mild cellular rejection when transplanted in the cornea of animals possessing natural antibody to a-gal epitope. The cornea reconstructed with porcine corneal stroma and HCEC had average cell density of 1721/mm^2 and had approximately 60% pump function compared with the normal cornea. These results indicated the possibility of clinical use of corneas reconstructed using HCEC.