Ex Vivo Expansion of human corneal endothelial cells for clinical application

    Ray Jui-Fang, Tsai MD
    Taipei Eye Center

    Purpose: To develop techniques to culture the corneal endothelial cells for clinical corneal endothelial keratoplasty. 

    Methods: Human donor corneas (from US Eye Bank) were selected for studies. Human corneal endothelium was obtained from the inner part of human limbal corneal rim. Human corneal endothelium was cut into a small pieces and cultured on the different kind of substrate including amniotic membrane. Series passages were done. Cultured human corneal endothelial cells finally were growth on the human corneal disc.

    Results: Age of human donor corneas was from 37 y/o to 70 y/o with mean 63+11 y/o. The death to cultured period was from 6 days to 10 days with mean 8.4 +1.8 days.  The cultured human corneal endothelial cells (HCEC) could be growth on the substrate with basement membrane substance. Serial passages from each HCEC were performed and results with 4.5 + 2.3 (2 to 9 passages) had been achieved from the aged human corneas.  We took one of p2 HCEC and cultured on the human corneal disc. About one month of culture the HCEC was confluent growth on the human corneal disc. TEM demonstrated mosaic pattern of HCEC on the cornea disc    
Conclusion: The ex vivo expansion of human corneal endothelial cells systems have been established. These techniques will allow us to further study the engineering of cornea.

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