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Marie-Claude Perron, Karine Zaniolo, Cristina Bostan, Olivier Rochette Drouin, Alexandre Deschambeault, Isabelle Brunette, Stéphanie Proulx; Successful Culture Of Human Corneal Endothelial Cells Isolated From Patients With Fuchs Endothelial Corneal Dystrophy. Invest. Ophthalmol. Vis. Sci. 2012;53(14):5988. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To assess the feasibility of culturing, without viral transduction, human corneal endothelial cells from patients with Fuchs endothelial dystrophy (FECD). We also evaluated which conditions yielded the best results in culture.
Descemet’s membranes (DM) excised from 29 consenting patients undergoing Descemet stripping automated endothelial keratoplasty (DSAEK) were used for this study. Endothelial cells were isolated and cultured. DM of 13 specimens were analyzed by transmission electron microscopy (TEM). Immunofluorescent staining for clusterin was also performed.
Of the 29 specimens, 18 successfully initiated a culture. Cell morphology varied from endothelial (rounded, slightly elongated cells, n=12) to fibroblastic-like (thin and very elongated cells, n=6). These differences in cell morphology were also observed with the normal human corneal endothelial cell cultures. Cultures initially presenting with an endothelial morphology maintained their shape in subculture. Similar levels of clusterin expression were observed in FECD and normal endothelial cells. TEM of FECD DM showed abnormalities typical of FECD, including a thickened DM, a posterior banded layer, a fibrillar layer and striated bodies of various sizes and periodicities. Patient’s preoperative corneal thickness, specimen size, presence of pigmentation and guttae were not predictive factors of culture success. Patient’s age however was correlated with culture success. Endothelial cells from patients younger than 60 years all generated successful cultures of endothelial morphology, while none of those from patients over 80 generated cultures of endothelial morphology. The absence of a fibrillar layer was also associated with greater success in culture.
This work shows for the first time that endothelial cells found on the central DM of patients suffering from FECD still retain proliferative capacity and that they can be isolated and cultured without viral transduction. This new in vitro model for FECD opens the way to the development of new treatments for this painful and blinding disease.
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