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Stephen Wahlig, Shu-Jun Lin, Khadijah Binte Adnan, Xin-Yi Seah, Matthew J Lovatt, Gary S L Peh, Jodhbir S Mehta; Optimization of Storage Conditions for Transportation of Cultured Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1375.
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© ARVO (1962-2015); The Authors (2016-present)
Cellular therapy for the treatment of corneal endothelial dysfunction is an emerging alternative to conventional allogeneic transplantation. The resources necessary for human corneal endothelial cell (HCEnC) propagation are primarily limited to major academic hospitals; however, therapeutic applications, on a broader scale, will require storage and delivery of cultured cells. The aim of this study was to determine optimal storage conditions for in vitro expanded corneal endothelial cells.
Primary human corneal endothelial cells, cultured using a dual media approach, were used to screen each storage condition. At 37°C, we tested the stabilization medium Human Endothelial-SFM (Endo-SFM) with and without serum, and an MEM-based organ culture medium. At room temperature (23°C) and 4°C we tested Endo-SFM and Optisol-GS. All plates were sealed with Parafilm to create a closed-storage system. After 48hrs in storage conditions, viability was determined with a Calcein AM (CAM) fluorescence microplate assay. Fluorescence CAM data was normalized using 37°C Endo-SFM as a reference.
There was no significant difference between Endo-SFM (1.0±0.03) and MEM (0.85±0.10) storage media at 37°C, or between Endo-SFM with and without serum at any temperature. Endo-SFM did perform significantly better than Optisol-GS at 23°C (0.77±0.10 v 0.48±0.02, p<0.01) and 4°C (1.05±0.03 v 0.25±0.04, p<0.001). Endo-SFM at 4°C also had significantly higher viability than at 23°C (p<0.01).
This data suggests that Endo-SFM stabilization medium best preserves cultured HCEnC viability at all storage temperatures, with optimal viability at 37°C and 4°C. Surprisingly Optisol-GS demonstrated poor viability in hypothermic conditions, despite its use for storage of whole corneas.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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