Comparison of different culture media for limbal epithelial cells cultivated ex vivo

• Grant number: 11/04596-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: August 01, 2011
• End date: April 30, 2013
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Mauro Nishi
• Grantee: Renata Ruoco Loureiro
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Several questions have been raised about the best technique for cultivation of limbal epithelial cells ex vivo. Variables such as culture medium, co-culture with feeder layer, substrate (scaffold) for culture must be well studied because they may influence survival and the maintenance of stemness of the cells to be transplanted. This study has been conducted to evaluate the effectiveness of different culture media on growth, proliferation, apoptosis and differentiation of limbal epithelial cells cultivated ex vivo. Corneal rims from different donors will have their posterior stroma removed and will be divided into 12 fragments and 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 will be statistically compared. (AU)