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P. Pagani, G. Campanile, A. Profumo, G. Bricola, C. Traverso; Fibrin–based substrate for in vitro cultivation as a platform for transplantation of Corneal Endothelial Cells. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4789.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To identify a biological substrate that would allow the proliferation of corneal endothelial cells cultured in vitro and the implication of the resulting construct in transplantation in vivo. Methods: Several substrates are evaluated and over then 100 primary cultures are established. In order to overcome the difficulties related to the manipulation of the tissue culture a film of fibrin was prepared obtained from a gel solution of fibrinogen and trombin pressed to obtain the minimum thickness compatible with the handling of the substrate. Human, bovine, porcine and rabbit corneal endothelial cells were isolated, plated and cultured up to confluence. The construct was layered above a human cornea previously deprived of endothelium; in this step, the construct was placed with the cultured cells facing the membrane of Descemet. Cornea and construct were incubated at 37°C for 2 days to allow the cultured cells to adhere to the cornea. Results:The fibrin film appeared a efficient substrate for cell adhesion, regardless the species used to harvest the endothelial cells. . With respect to proliferation, the cells obtained from bovine, porcine and rabbit cornea were readily growing, reaching confluence within 5 days. The cells of human origin presented a lower proliferation rate, requiring more than 3 weeks to reach confluence. During incubation, all the cornea constructs presented some stromal edema. This is routinely observed during corneal preservation. Hystology revealed that the endothelial cells grew on the fibrin film adhered to the Descemet’s membrane. The experiments of in vivo transplantation in rabbit are ongoing. Conclusions: Our work shows the efficiency of this method here developed, based on the use of a fibrin film to allow endothelial cells proliferation. The composite cells/film can be detached from the dish as a whole and directly implanted in a host eye in a rabbit model.
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