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T Kim, EP Kay, RE Smith; Growth Characteristics of Human and Rabbit Corneal Endothelial Cells on Various Matrices . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3186.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine the role of various matrices on primary human and passaged rabbit corneal endothelial cells (CEC) culture. Methods: Human CEC, prepared from donor corneal rims after corneal transplantation, and passaged rabbit CEC were seeded on various matrices including preserved human amniotic membrane, extracellular matrix (ECM) secreted by rabbit CEC, rabbit Descemet`s membrane, and rabbit whole cornea. The regular culture plate served as a control. Phase contrast microscopy and vital staining were done to compare growth characteristics. Results: When human and rabbit CEC were seeded on human amniotic membrane with intact amniotic epithelium, clumps of cells formed and did not spread; but without amniotic epithelium, cells adhered evenly and proliferation occurred at the same speed observed in controls. After removal of preexisting endothelium with 0.14% ammonium hydroxide on rabbit ECM and Descemet`s membrane, human and rabbit cells were found to adhere and grow faster than those on the controls. The subculture from CEC on rabbit ECM and Descemet`s membrane had the more proliferative capacity than those on control plate. Vital staining of passaged rabbit CEC cultured on endothelium-removed rabbit whole cornea showed faint intercellular outlines; with detachment of its Descemet`s membrane and examination by phase contrast microscopy, good visualization of cultured CEC could be obtained. Conclusion: These data suggest that ECM plays a role in adhesion and proliferation of human and rabbit CEC; and denuded human amniotic membrane and rabbit Descemet`s membrane are good substrates for cultivation of CEC.
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