Improved preparation and preservation of human keratoplasty lenticules

Abstract

Purpose: To improve the preparation of lenticules from human cornea and to obtain their preservation without loss of viable keratocytes.

Methods: The epithelium was manually removed after bathing the surface of the cornea with a solution of trypsin and EDTA. Lenticules were prepared by microkeratome resection and viable keratocytes were visualized by staining with thiazolyl blue (MTT).

Results: The pretreatment with trypsin-EDTA allowed the removal of the epithelium without damage to the keratocytes and the stroma. When these lenticules were incubated in Optisol-GS for 7 days at 4 degrees C, they showed a limited thickness increase and a preservation of keratocyte viability.

Conclusion: This procedure allows the preparation of lenticules with viable keratocytes that can be preserved in the cold for at least 1 week.