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Increased Endothelial Survival of Organ-Cultured Corneas Stored in FGF-2–Supplemented Serum-Free Medium

¹From the Department of Ophthalmology, Charité Medical Faculty, Humboldt University Berlin, Germany; and the ²Department of Ophthalmology, Geneva University Hospital, Geneva, Switzerland.

PURPOSE. To investigate the effect of FGF-2 on corneal endothelial cell survival in porcine and human corneas during corneal storage in a serum-free medium.

METHODS. Porcine and paired human corneas were stored at 32°C for 9 and 22 days, respectively. One cornea of each pair was stored in a serum-free culture medium, and the mate was preserved in the same medium supplemented with 10 ng/mL FGF-2. Quantitative analysis of corneal damage after storage was determined by the Janus green photometry technique. 5-Bromo-2-deoxyuridine (BrdU) labeling of the endothelium determined the effect of FGF-2 on endothelial proliferation during storage. Additional cell culture studies were performed to elucidate the role of FGF-2 on the incidence of endothelial apoptosis after serum deprivation.

RESULTS. When FGF-2 was added to the serum-free medium, the damage rates of porcine endothelia were reduced from 15.1% ± 8.7% (control) to 6.4% ± 2.0% after 9 days and from 25.3% ± 10.2% to 13.6% ± 4.2% after 22 days of storage. In the human corneas stored during 22 days in FGF-2–supplemented medium, the amount of endothelial damage was 11.8% ± 3.2%, which was significantly less damage than in the control fellow corneas stored in unsupplemented serum-free medium (19.3% ± 6.3%; P < 0.01). DNA synthesis was not enhanced in corneas stored in serum-free medium, serum-free medium+FGF-2, or medium containing 10% FCS. Only a few (3.8%) TUNEL-positive endothelial cells were detected in cultures maintained in FGF-2–supplemented serum-free medium compared with a high number (48%) of apoptotic cells in control cultures.

CONCLUSIONS. FGF-2 efficiently reduces human corneal endothelial damage that occurs during organ culture storage in a serum-free medium. This effect is truly protective, because no proliferative activity and a decreased rate of apoptosis were determined. FGF-2 emerges as an important component of a future serum-free corneal organ-culture medium established to replace fetal calf serum (FCS) as a potential source of recipient infection.

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