Abstract
Purpose :
Human platelet lysate (hPL) as a replacement for fotal bovine serum (FBS) in culturing human corneal endothelium is an emerging area of interest. We previously published research results showing that in vitro hPL can be considered as a supplement to replace FBS in human corneal endothelial cell culture. This study is to further evaluate hPL as a replacement for FBS in Eye Bank organ culture medium (OCM) to preserve human donor corneas.
Methods :
Six pairs of human corneas were obtained from New South Wales Tissue Bank. To minimise the variation, all right corneas from the pairs were allocated in FBS supplement medium and left corneas were in hPL medium. Cornea assessment and preservation were followed by standardized Eye Bank procedures for processing and storage of organ culture corneas. Corneal endothelial cell (CEC) counts and viability tests were performed at Days 0, 7, 14 and 21. Expression and localization of Na+/K+-ATPase, ZO-1 and CD166 on corneas stored in hPL and FBS media were assessed with immunofluorescence and immunoblotting at Day 21. Paired t-tests were used to compare cell counts and viabilities between two groups. P<0.05 was considered as statistical significance.
Results :
Six pairs of corneas from 5 male and 1 female completed the day 21 preservation. The mean age of the donors was 65.2 years old (range 49-93). The cause of death for the donors were malignancy (33.3%), a cardiac event (33.3%) and a cerebral event (33.3%). The CEC counts (mean±SD) in FBS supplement media at Day 0, 7, 14 and 21 were 3014.72±1162.23, 2860.47±1107.64, 2628.67±1024.43 and 2348.67±894.81 cells/mm2 respectively, compared to 2946.73±310.05, 2945.74±395.26, 2601.49±153.23 and 2303.79±157.08 cells/mm2 in hPL supplement media (all p>0.05). The cell viability values as percentage were 96.8±1.6, 96.8±1.3, 94±2.0 and 91.6±3.85 in FBS supplement media and 96.8±1.6, 96.2±2.0, 95.2±1.1 and 86±8.03 in hPL supplement media at Day 0, 7, 14 and 21, respectively (all p>0.05). No differences in expression and localization of Na+/K+-ATPase, ZO-1 and CD166 were observed between corneas stored in hPL and FBS supplement media.
Conclusions :
There was no difference in endothelial cell counts, cell viability and immune mark expressions at all storage stages between hPL and FBS supplement OCM. hPL is at least equivalent to FBS for human corneal storage but further studies are needed to confirm its effectiveness.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.