Culture of human corneal endothelial cells isolated from corneas with Fuchs endothelial corneal dystrophy

The purpose of this study was to assess the feasibility of initiating primary cultures of corneal endothelial cells from patients suffering from Fuchs endothelial corneal dystrophy (FECD; MIM# 1036800). We also evaluated which conditions yielded the best results for culture. Twenty-nine patients und...

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Veröffentlicht in:Experimental eye research 2012, Vol.94 (1), p.22-31
Hauptverfasser: Zaniolo, Karine, Bostan, Cristina, Rochette Drouin, Olivier, Deschambeault, Alexandre, Perron, Marie-Claude, Brunette, Isabelle, Proulx, Stéphanie
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container_end_page 31
container_issue 1
container_start_page 22
container_title Experimental eye research
container_volume 94
creator Zaniolo, Karine
Bostan, Cristina
Rochette Drouin, Olivier
Deschambeault, Alexandre
Perron, Marie-Claude
Brunette, Isabelle
Proulx, Stéphanie
description The purpose of this study was to assess the feasibility of initiating primary cultures of corneal endothelial cells from patients suffering from Fuchs endothelial corneal dystrophy (FECD; MIM# 1036800). We also evaluated which conditions yielded the best results for culture. Twenty-nine patients undergoing Descemet stripping automated endothelial keratoplasty consented to the use of their excised Descemet's membrane for this study. Out of the 29 specimens, 18 successfully initiated a culture. Cell morphology varied between endothelial (rounded, slightly elongated cells, n = 12) and 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. The cultures that initially presented an endothelial morphology maintained their shape in subcultures. Clusterin expression was similar in FECD and normal endothelial cells. Transmission electron microscopy of FECD Descemet's membranes showed a high degree of various abnormalities generally found in this disease, such as a thickened Descemet's membrane, presence of a posterior banded layer, presence of a fibrillar layer and striated bodies of various sizes and periodicities. Patient's age was predictive of culture success, all younger FECD donors generating cultures of endothelial morphology. The absence of a fibrillar layer was also a factor associated with greater success. Culture success was not dependent on specimen size, specimen pigmentation, or patient's preoperative central corneal thickness. In conclusion, this paper shows for the first time that central Descemet's membranes of patients suffering from FECD possess proliferative endothelial cells that can be isolated and cultured without viral transduction, opening the way for new in vitro studies of this disease. ► We cultured corneal endothelial cells from Fuchs corneal endothelial dystrophy (FECD) specimens. ► Sixty-two percent of FECD specimens successfully generated a culture. ► FECD endothelial cells adopted both endothelial and fibroblastic-like morphologies in culture. ► Patient's age and absence of a fibrillar layer were factors that improved culture success.
doi_str_mv 10.1016/j.exer.2011.10.018
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We also evaluated which conditions yielded the best results for culture. Twenty-nine patients undergoing Descemet stripping automated endothelial keratoplasty consented to the use of their excised Descemet's membrane for this study. Out of the 29 specimens, 18 successfully initiated a culture. Cell morphology varied between endothelial (rounded, slightly elongated cells, n = 12) and 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. The cultures that initially presented an endothelial morphology maintained their shape in subcultures. Clusterin expression was similar in FECD and normal endothelial cells. Transmission electron microscopy of FECD Descemet's membranes showed a high degree of various abnormalities generally found in this disease, such as a thickened Descemet's membrane, presence of a posterior banded layer, presence of a fibrillar layer and striated bodies of various sizes and periodicities. Patient's age was predictive of culture success, all younger FECD donors generating cultures of endothelial morphology. The absence of a fibrillar layer was also a factor associated with greater success. Culture success was not dependent on specimen size, specimen pigmentation, or patient's preoperative central corneal thickness. 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Transmission electron microscopy of FECD Descemet's membranes showed a high degree of various abnormalities generally found in this disease, such as a thickened Descemet's membrane, presence of a posterior banded layer, presence of a fibrillar layer and striated bodies of various sizes and periodicities. Patient's age was predictive of culture success, all younger FECD donors generating cultures of endothelial morphology. The absence of a fibrillar layer was also a factor associated with greater success. Culture success was not dependent on specimen size, specimen pigmentation, or patient's preoperative central corneal thickness. 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MIM# 1036800). We also evaluated which conditions yielded the best results for culture. Twenty-nine patients undergoing Descemet stripping automated endothelial keratoplasty consented to the use of their excised Descemet's membrane for this study. Out of the 29 specimens, 18 successfully initiated a culture. Cell morphology varied between endothelial (rounded, slightly elongated cells, n = 12) and 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. The cultures that initially presented an endothelial morphology maintained their shape in subcultures. Clusterin expression was similar in FECD and normal endothelial cells. Transmission electron microscopy of FECD Descemet's membranes showed a high degree of various abnormalities generally found in this disease, such as a thickened Descemet's membrane, presence of a posterior banded layer, presence of a fibrillar layer and striated bodies of various sizes and periodicities. Patient's age was predictive of culture success, all younger FECD donors generating cultures of endothelial morphology. The absence of a fibrillar layer was also a factor associated with greater success. Culture success was not dependent on specimen size, specimen pigmentation, or patient's preoperative central corneal thickness. In conclusion, this paper shows for the first time that central Descemet's membranes of patients suffering from FECD possess proliferative endothelial cells that can be isolated and cultured without viral transduction, opening the way for new in vitro studies of this disease. ► We cultured corneal endothelial cells from Fuchs corneal endothelial dystrophy (FECD) specimens. ► Sixty-two percent of FECD specimens successfully generated a culture. ► FECD endothelial cells adopted both endothelial and fibroblastic-like morphologies in culture. ► Patient's age and absence of a fibrillar layer were factors that improved culture success.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>22134119</pmid><doi>10.1016/j.exer.2011.10.018</doi><tpages>10</tpages></addata></record>
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subjects Aged
Aged, 80 and over
Aging - physiology
Biomarkers - metabolism
cell culture
Cell Culture Techniques
Cell Proliferation
Cell Separation
Cell Shape
Clusterin - metabolism
cornea guttata
corneal endothelial cells
Descemet Membrane - ultrastructure
Descemet's membrane
Endothelium, Corneal - metabolism
Endothelium, Corneal - pathology
Female
Fluorescent Antibody Technique, Indirect
Fuchs endothelial corneal dystrophy
Fuchs' Endothelial Dystrophy - metabolism
Fuchs' Endothelial Dystrophy - pathology
Humans
Keratins - metabolism
Male
Middle Aged
title Culture of human corneal endothelial cells isolated from corneas with Fuchs endothelial corneal dystrophy
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