Conjunctival fibroblasts were derived from normal human conjunctivas that were obtained from individuals undergoing eyelid or strabismus surgery. All the cells were positive for vimentin and negative for cytokeratin, suggesting no contamination of the cultures with epithelial cells. No changes in cell morphology or immunoreactivity were apparent during culture for up to 8 passages.
Two normal human conjunctivas were obtained upon informed consent of individuals undergoing eyelid or strabismus surgery, and were processed separately.
Each conjunctiva was incubated for 60 min at 370C with dispase (2 mg/ml), and epithelial cells were mechanically removed. The remaining tissue was digested with collagenase (2 mg/ml), for 1 hr at 370C. The cells from each conjunctiva were cultured separately in MEM supplemented with fetal bovine serum (10%).