Cultivated autologous limbal epithelial cells (CALEC): product development, manufacture, and initial evaluation of feasibility

Abstract: To treat unilateral limbal stem cell (LSC) deficiency, we developed a novel cell therapy: cultivated autologous limbal epithelial cells (CALEC). The manufacturing platform employed reproducible methods for LSC isolation and expansion onto human amniotic membrane and rigorous quality control measures without use of feeder cells or serum. Small limbal biopsies were used to generate CALEC constructs in an average of 16 days. Cultivated cells maintained epithelial cell phenotype with colony forming and proliferative capacities, with high viability and metabolic activity during manufacture and transportation. Analysis of LSC biomarkers showed preservation of ‘stemness’ in CALEC constructs. After pre-clinical development, CALEC grafts were manufactured in a current good manufacturing practices compliant facility. A Phase I clinical trial enrolled five patients with unilateral LSC deficiency. Clinical case histories with 12 months follow-up are reported for the first four patients who received successful CALEC transplants. This initial clinical trial establishes feasibility of this technique with no immediate safety concerns.