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Growth Factors as a Supplement to Serum-free Culture-media for Corneal Organ Culture An Experimental Study
1Rieck P., 2Gigon M., 1Jaeckel C., 1Metzner S., 1Hart K., 1Hartmann C.,
1Humboldt-Universität zu Berlin, Charité Campus Virchow-Klinikum, Augenklinik (Berlin)
2Service d´Ophtalmologie, CHU Genève (Genf)
Purpose: To investigate the effect of FGF-2 on corneal endothelial cell damage in porcine and human corneas during corneal storage.
Methods: Porcine and paired human corneas were stored at 32°C for 9 resp. 22 days. 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. Quantitation of corneal damage after storage was determined using the Janus green photometry technique as well as light microscopical and ultrastructural investigations. The effect of FGF-2 on endothelial proliferation during storage was investigated by means of BrdU-labeling as an indictor of DNA synthesis.
Results: The porcine corneas preserved in serum free medium revealed a mean overall endothelial damage of 15.1 ± 8.7% after 9 days and 25.3 ± 10.2% after 22 days of storage. When FGF-2 was added to the medium the damage rates were reduced to 6.4 ± 2.0% after 9 days and to 15.6 ± 4.2% after 22 days of storage. The human corneas preserved in serum free medium for 22 days revealed a mean overall endothelial damage of 19.3 ± 6.3%. In the mate corneas stored in FGF-2 supplemented medium, the amount of endothelial damage was 11.8 ± 3.2% (p<0.01). Light- and electron microscopical investigations confirmed this quantitative determination of a protective effect of FGF-2 on the human endothelium. DNA synthesis was not enhanced in corneas stored in serum free medium, serum free medium + FGF-2 or medium containing 10% FCS, respectively.
Conclusions: This study demonstrates the effectiveness of FGF-2 to protect human corneal endothelium from damage occuring during organ culture storage in a serum free medium. This effect is truely protective since no proliferative activity could be determined. FGF-2 could be a candidate to replace the widespread use of fetal calf serum which remains a potential source of infection, especially of prion diseases.