A New Technique for Harvesting Limbal Stem Cell Tissue for Transplantation Using an Automated Microkeratome and a Novel Globe-Fixation System.

Abstract

To introduce an unprecedented technique, instrumentation, and setup for the superficial limbus harvest from the human cadaver donor whole globe. We studied several superficial limbus harvest alternatives, developed a preferred approach with our available instruments, and optimized it on a handful of (seropositive) cadaver donors of whole globes. The globe was pressurized to about normal intraocular pressure by viscoelastic injection through the optic nerve stump. The globe was then mounted on a dynamic globe fixator that maintained a negative pressure in its stabilizing socket. Exertion of the negative pressure effectively elevated globe intraocular pressure (to over 40 mm Hg) and made the corneoscleral wall tight. The socket was then held tilted to the left side for about 35 to 40 degrees to put the limbal zone horizontally and mildly "chin-up." Next, the microkeratome was put on the uppermost and exposed limbus of the globe and activated, and the socket was rotated clockwise under the microkeratome head and its oscillating blade, allowing an effective 360-degree revolution of the microkeratome head around the limbal belt (for a right-handed operator and a counterclockwise cut). We consistently succeeded in peeling intact 360-degree strips of the smooth superficial limbus by using blades with varying depths. Our method can be further equipped and optimized and be used by the eye banks and the surgeons for keratolimbal grafting as a more efficient limbal stem cell tissue harvest technique.