Human Anterior Lens Capsule as a Biologic Substrate for the Ex Vivo Expansion of Limbal Stem Cells in Ocular Surface Reconstruction

Abstract

To study the potential use of human anterior lens capsule as a scaffold for stem cell transplantation in treatment of limbal cell deficiency. Limbal biopsies and anterior lens capsules were obtained (same eye) from 30 patients during cataract surgery. Biopsies were suspended in Dulbecco modified Eagle medium under sterile conditions and stored at 4 degrees C. Capsules were treated in distilled water under strict asepsis for 2 hours to eliminate the crystalline epithelium and stored at 4 degrees C. After initial processing, the limbal biopsy was plated epithelial-side down (48 hours) on the capsular specimen in a 35-mm culture dish. Samples were sorted into 4 groups. Group 1 was made up of 10 samples in which limbal biopsies were allowed to grow on corresponding capsules from the same eye (autologous). Group 2 was 10 limbal biopsies that were allowed to grow on capsules of different eye (allogeneic). The remaining specimens were randomized into 2 groups. Group 3 included 10 capsules on which an ex vivo expanded cell line was allowed to grow. Group 4 harbored 10 limbal biopsies that were allowed to grow on polystyrene culture plates. All specimens were incubated for 2 weeks at 37 degrees C and 5% CO2. Cell density, viability, morphology, and adherence of the cell-capsule complex were evaluated at 1, 3, 7, and 14 days. Rate of cell growth and density in groups 1 and 2 were comparable to the control groups. Cell viability was 95% or superior in all groups, and desmosomes developed between growing cells. Human anterior lens capsule is a potential scaffold for ex vivo expansion of limbal epithelial cells, possibly providing a substrate for ocular surface reconstruction.