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P32-A134 Cytoprotective effects of human platelet lysate during the xeno-free culture of human donor corneas
  1. Delia Talpan1,
  2. Sabine Salla1,2,
  3. Linus Meusel1,2,
  4. Peter Walter1,2,
  5. Chao-Chung Kuo3,
  6. Julia Franzen3,
  7. Matthias Fuest1,2
  1. 1Department of Ophthalmology, RWTH Aachen University, Aachen, Germany
  2. 2Cornea Bank Aachen, RWTH Aachen University, Aachen, Germany
  3. 3Institute for Interdisciplinary Center for Clinical Research (IZKF), RWTH Aachen University, Aachen, Germany


Purpose We evaluated the suitability of 2% human platelet lysate (2%HPL) to replace 2% fetal bovine serum containing medium (2%FBS) for the xeno-free organ culture of human donor corneas.

Methods 32 human corneas unsuitable for transplantation from 16 human donors (age 69.3±15.7years) were collected 38.5±17.1 hours after death. They were first cultured in 2%FBS containing medium for 3 days (time point TP1), then evaluated by phase contrast microscopy (endothelial cell density (ECD) and cell morphology. Following an additional 25-days culture period (time point TP2) in either 2%FBS or 2%HPL medium the pairs were again compared by phase contrast microscopy (ECD and morphology), stroma and Descemet membrane/endothelium (DmE) were processed for next generation sequencing (NGS).

Results ECD did not differ between the 2%HPL and 2%FBS group at TP1 (p=0.87). At TP2 the ECD was higher in the 2%HPL group (2179±288cells/mm2) compared to 2%FBS (2113±331cells/mm2; p=0.03), and endothelial cell loss was lower (ECL hPL=-0.7% vs. FBS=-3.8%; p=0.01). There were no significant differences in cell morphology, neither between TP1 and 2 nor between 2%HPL and 2%FBS. NGS showed the differential expression of 1644 genes in endothelial and 217 genes in stromal cells. 2%HPL led to the upregulation of cytoprotective, anti-inflammatory and anti-fibrotic genes (e.g. HMOX1, SERPINE1, ANGPTL4, LEFTY2, GADD45B, PLIN2, PTX3, GFRA1/2) and the downregulation of pro-inflammatory/apoptotic genes (e.g. CXCL14, SIK1B, PLK5, PPP2R3B, SLURP1, FABP5, MAL, GATA3).

Conclusion 2%HPL is a suitable xeno-free substitution for 2%FBS in human cornea organ culture, inducing less ECL and potentially beneficial alterations in gene expression.

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