Serum‐free cornea culture with hydroxyethyl starch as a deswelling agent

Abstract

AbstractPurpose Varying states of hydration during organ culture and subsequent dehydration of corneas with Dextran prior to transplantation cause endothelial cell damage. Hydroxyethyl starch 130 (HES) has been suggested as a permanent dehydrating agent in serum‐free cornea culture. This study compares corneas stored in a synthetic medium containing HES with corneas stored and dehydrated under standard conditions.Methods Twenty pairs of human donor corneas were cultivated in MEM with antibiotics at 31°C for 16 days (groups A and B) or 28 days (groups C and D, n=10). Media contained 2%FCS in group A and C, and 7.5 % HES 130 plus 1mU/l human insulin in groups B and D. On day 15 or 27, corneas from group A and C were placed in dehydration medium (MEM + antibiotics + 2%FCS + 5% dextran 500). Endothelial cell density and morphology were investigated at the beginning and end of culture. Corneal thickness was assessed by pachymetry. Keratocyte viability was assessed by TUNEL and Annexin V staining.Results Endothelial cell loss was higher in group A (278.5+/‐184.2) vs. B (156.6+/‐154; p=0.0293), and in group C (490+/‐156.3) vs. D (301.8+/‐189.6; p=0.0195), with similar morphology. Final corneal thickness was 882+119µm and 914+119µm in groups A and B, and 889+99µm and 957+87µm in groups C and D, resp. (n.s.). No significant differences were found in TUNEL and Annexin staining.Conclusion The synthetic medium supplemented with HES 130 and insulin improves endothelial cell survival during cornea culture, without adverse effects ion keratocyte viability. It is thus feasible to store donor corneas in fully synthetic medium without the chemically undefined FCS and without the need for dehydration, resulting in increased safety and faster transplant availability.