Human anterior lens capsule as a biologic substrate for the ex vivo expansion of limbal stem cells in ocular surface reconstruction

Purpose: To study the potential use of human anterior lens capsule as a scaffold for stem cell transplantation in treatment of limbal cell deficiency. Methods: 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 I 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. Results: 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. Conclusions: Human anterior lens capsule is a potential scaffold for ex vivo expansion of limbal epithelial cells, possibly providing a substrate for ocular surface reconstruction.