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Y. Ishino, Y. Sano, S. Kinoshita; Survival of Transplanted Cultivated Human Corneal Endotheial Cells In Vivo . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4330.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: We reported the feasibility of amniotic membrane (AM) as a carrier for cultivated human corneal endothelial cells (cHCECs) and the function of cHCECs in vivo (2002, ARVO). We wished to examine whether cHCECs transplanted in vivo survive after transplantation. Methods: Corneal tissue (47 years old, male, endothelial cell density: 3065cells/mm2) was obtained from U.S. eye bank. To grow HCECs, Descemet's membrane with HCECs was dissected from stroma. HCECs were released by dispase treatment, and cell suspension was seeded in a well of 24 well plate. After 5 passages, cHCECs were labeled with fluorescent membrane dye DiI (DiIC18 (3)), and seeded on AM that denuded of epithelial cells by scraping. After 2 weeks cultivation, cHCEC sheets using AM as a carrier were transplanted onto rabbits cornea in vivo. Examination of corneal appearance via slit-lamp biomicroscopy and ultrasonic pachymetery were procedured, up to Day 7 after operation daily, up to Day 28 weekly. At 7 days and 28 days after operation, we sacrificed these animals, removed the grafts, and observed them under the fluorescence microscope. Results: At both 7 days and 28 days after operation, control grafts consisting of transplanted only acellular AM after stripping Descemet's membrane became highly edematous (Corneal thickness; Day 7 after operation: 974µm, Day28: over 1000µm), neverthless grafts with cHCECs on AM had little edema and excellent transparency (Corneal thickness Day 7 after operation: 411µm, Day28: over 506µm). Transplanted cHCECs labeled with DiI were survived with localizing on AM (cell density; Day 7 after operation: 2432cells/mm2, Day28: over 1015 cells/mm2), and non-labeled rabbit corneal endothelial cells were not invaded. Conclusions: Transplanted cHCECs with using AM as its carrier had survived and functioned in vivo till day 28 after operation.
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