Abstract
Purpose: :
Evaluate the effectiveness of different culture media on growth, proliferation, apoptosis and differentiation of limbal epithelial cells cultivated ex vivo.
Methods: :
Corneal rims from different donors had their posterior stroma removed and were cultured in three different culture media: SHEM, KSFM, Epilife. The epithelial cell cultures were submitted to analysis of growth and epithelial migration; immunocytochemistry for ABCG2, p63, Ki67, CK3 and VMT; RT-PCR; and cell viability test with Hoechst. All results were statistically compared.
Results: :
Epithelial cells cultivated in SHEM medium presented rapid and progressive growth, with a highly positive percentage of cells expressing epithelial cytokeratin CK3. The epithelial cells cultivated in KSFM showed epithelial and mesenchymal appearance and high positivity for ABCG2, p63, Ki67 and VMT. A similar pattern of antigen expression was noted with the epithelial cells cultivated in Epilife media. However, in the latter, cells presented only an epithelial phenotype.
Conclusions: :
We conclude that Epilife and KSFM seem to be the best media for establishing limbal epithelial cell cultures. They contain low calcium concentration and keep the cells in a more undifferentiated status when compared to cells cultured in SHEM medium.
Keywords: cornea: basic science • cornea: stroma and keratocytes