Abstract
Purpose:
The purpose of this study was to assess the feasibility of initiating primary cultures of corneal endothelial cells from patients suffering from various corneal diseases. We also evaluated which conditions yielded the best results for culture.
Methods:
Consenting patients undergoing penetrating keratoplasty or Descemet’s stripping automated endothelial keratoplasty were enrolled in this study. The cornea (or Descemet’s membrane), removed at the time of surgery, was sent to the laboratory. Upon receipt, specimens were photographed, endothelial cells were isolated and cultured, and Descemet’s membranes were fixed in 3.7% formaldehyde for histology. Data collected included diagnosis, age and sex of the donor, time spent in Optisol, culture success and number of cells obtained after a primary culture. Control healthy Eye Bank corneas were processed in a similar manner.
Results:
A total of 194 specimens were obtained between August 2009 and November 2012. Pathologies included Fuchs corneal endothelial dystrophy (FECD; n=93), pseudophakic bullous keratoplathy (PBK; n=33), FECD+PBK (n=4) and other corneal disorders (n=64). Overall, 56 of the 194 diseased specimens (FECD n=37; PBK n=3; FECD+PBK n=2; other n=14) and all of the 29 healthy corneas successfully initiated an endothelial cell culture. Among the diseased specimens, the mean(±SD) donor age was 66.5 ±11.9 years for successful cultures and 68.9 ±15.2 years for the unsuccessful cultures. Time spent in Optisol was 2.7 ±1.2 and 3.2 ±2.6 days for successful and unsuccessful cultures, respectively. The diseased specimens yielded 120 000 ±86 000 cells and the healthy corneas 122 000 ±55 000 cells.
Conclusions:
This study shows that successful corneal endothelial cell culture can be generated despite various corneal diseases. FECD allowed the highest success rate. Once culture was initiated, a similar number of cells was obtained from FECD, BPK and healthy specimens. Patient sex and age, and the time spent by the specimen in Optisol did not influence success rate. The Quebec Corneal Cell Bank will become a useful tool for the study of various corneal endotheliopathies.
Keywords: 481 cornea: endothelium •
638 pathology: human