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JA Dixon, JM Prince, I Reynolds, AB Tullo, MC Hillarby; Changes in Gene Expression During Organ Culture of the Human Cornea . Invest. Ophthalmol. Vis. Sci. 2002;43(13):133.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Prior to transplantation corneas are stored in organ culture for up 4 weeks. However, up to 30% of corneas are discarded, mainly due to a poor endothelial cell count. An understanding of the mechanisms, which may be involved in endothelial cell loss could lead to improved culture conditions which would increase the number of corneas available for transplantation. An investigation into genes regulation during the culture period prior to transplantation, was therefore undertaken. Methods: RNA was extracted from different corneas at six time points during organ culture (days: 0, 3, 7, 13, 21 and 29). Following RT-PCR using the Clontech BD's SMART cDNA synthesis method, radioactive micro array analysis was performed using individual apoptosis Atlas arrays for each time point and analysed using Atlas Image 1.5 software. Results: A number of genes involved in signalling pathways (RhoA, IGFBP-6, GADD153), cell cycling (cdks & cyclins, CAK, cdcs, Retinoblastoma, c-myc) and apoptosis (Caspases, TNF-RI, TNF-RII, Fas/Apo1, Clusterin, PIG3, Bad, RATS-1, DAD-1) were found to be upregulated at certain time points during the culture period. Conclusion: The group of genes upregulated provides further evidence for apoptosis of corneal cells during organ culture, which may involve the TNF-α induced pathway. Immunohistochemistry is being performed to confirm the localisation of the expressed proteins.
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