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 |
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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 |
format | Article |
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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.</description><identifier>ISSN: 0014-4835</identifier><identifier>EISSN: 1096-0007</identifier><identifier>DOI: 10.1016/j.exer.2011.10.018</identifier><identifier>PMID: 22134119</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>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</subject><ispartof>Experimental eye research, 2012, Vol.94 (1), p.22-31</ispartof><rights>2011 Elsevier Ltd</rights><rights>Copyright © 2011 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c355t-aec43373dbefc021534dac6b15592ab87afc1bbc03d1758bfeaab821ef6a868f3</citedby><cites>FETCH-LOGICAL-c355t-aec43373dbefc021534dac6b15592ab87afc1bbc03d1758bfeaab821ef6a868f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0014483511003344$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,4010,27900,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22134119$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zaniolo, Karine</creatorcontrib><creatorcontrib>Bostan, Cristina</creatorcontrib><creatorcontrib>Rochette Drouin, Olivier</creatorcontrib><creatorcontrib>Deschambeault, Alexandre</creatorcontrib><creatorcontrib>Perron, Marie-Claude</creatorcontrib><creatorcontrib>Brunette, Isabelle</creatorcontrib><creatorcontrib>Proulx, Stéphanie</creatorcontrib><title>Culture of human corneal endothelial cells isolated from corneas with Fuchs endothelial corneal dystrophy</title><title>Experimental eye research</title><addtitle>Exp Eye Res</addtitle><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.</description><subject>Aged</subject><subject>Aged, 80 and over</subject><subject>Aging - physiology</subject><subject>Biomarkers - metabolism</subject><subject>cell culture</subject><subject>Cell Culture Techniques</subject><subject>Cell Proliferation</subject><subject>Cell Separation</subject><subject>Cell Shape</subject><subject>Clusterin - metabolism</subject><subject>cornea guttata</subject><subject>corneal endothelial cells</subject><subject>Descemet Membrane - ultrastructure</subject><subject>Descemet's membrane</subject><subject>Endothelium, Corneal - metabolism</subject><subject>Endothelium, Corneal - pathology</subject><subject>Female</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Fuchs endothelial corneal dystrophy</subject><subject>Fuchs' Endothelial Dystrophy - metabolism</subject><subject>Fuchs' Endothelial Dystrophy - pathology</subject><subject>Humans</subject><subject>Keratins - metabolism</subject><subject>Male</subject><subject>Middle Aged</subject><issn>0014-4835</issn><issn>1096-0007</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9LxDAQxYMouq5-AQ_Sm6fWTNN_C15kcVVY8KLnkCYTmqVt1qRV99ubsqvgxdMMj_feMD9CroAmQKG43ST4hS5JKUAQEgrVEZkBXRQxpbQ8JjNKIYuziuVn5Nz7TVBZVman5CxNgWUAixkxy7EdRoeR1VEzdqKPpHU9ijbCXtmhwdaEXWLb-sh424oBVaSd7Q4-H32aoYlWo2z838ihRu384Oy22V2QEy1aj5eHOSdvq4fX5VO8fnl8Xt6vY8nyfIgFyoyxkqkataQp5CxTQhY15PkiFXVVCi2hriVlCsq8qjWKoKaAuhBVUWk2Jzf73q2z7yP6gXfGTw-IHu3o-QKKHKosHJmTdO-UznrvUPOtM51wOw6UT4T5hk-E-UR40gLhELo-1I91h-o38oM0GO72BgxPfpgQ99JgL1EZh3Lgypr_-r8BllKPvA</recordid><startdate>2012</startdate><enddate>2012</enddate><creator>Zaniolo, Karine</creator><creator>Bostan, Cristina</creator><creator>Rochette Drouin, Olivier</creator><creator>Deschambeault, Alexandre</creator><creator>Perron, Marie-Claude</creator><creator>Brunette, Isabelle</creator><creator>Proulx, Stéphanie</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>2012</creationdate><title>Culture of human corneal endothelial cells isolated from corneas with Fuchs endothelial corneal dystrophy</title><author>Zaniolo, Karine ; Bostan, Cristina ; Rochette Drouin, Olivier ; Deschambeault, Alexandre ; Perron, Marie-Claude ; Brunette, Isabelle ; Proulx, Stéphanie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c355t-aec43373dbefc021534dac6b15592ab87afc1bbc03d1758bfeaab821ef6a868f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Aged</topic><topic>Aged, 80 and over</topic><topic>Aging - physiology</topic><topic>Biomarkers - metabolism</topic><topic>cell culture</topic><topic>Cell Culture Techniques</topic><topic>Cell Proliferation</topic><topic>Cell Separation</topic><topic>Cell Shape</topic><topic>Clusterin - metabolism</topic><topic>cornea guttata</topic><topic>corneal endothelial cells</topic><topic>Descemet Membrane - ultrastructure</topic><topic>Descemet's membrane</topic><topic>Endothelium, Corneal - metabolism</topic><topic>Endothelium, Corneal - pathology</topic><topic>Female</topic><topic>Fluorescent Antibody Technique, Indirect</topic><topic>Fuchs endothelial corneal dystrophy</topic><topic>Fuchs' Endothelial Dystrophy - metabolism</topic><topic>Fuchs' Endothelial Dystrophy - pathology</topic><topic>Humans</topic><topic>Keratins - metabolism</topic><topic>Male</topic><topic>Middle Aged</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zaniolo, Karine</creatorcontrib><creatorcontrib>Bostan, Cristina</creatorcontrib><creatorcontrib>Rochette Drouin, Olivier</creatorcontrib><creatorcontrib>Deschambeault, Alexandre</creatorcontrib><creatorcontrib>Perron, Marie-Claude</creatorcontrib><creatorcontrib>Brunette, Isabelle</creatorcontrib><creatorcontrib>Proulx, Stéphanie</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Experimental eye research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zaniolo, Karine</au><au>Bostan, Cristina</au><au>Rochette Drouin, Olivier</au><au>Deschambeault, Alexandre</au><au>Perron, Marie-Claude</au><au>Brunette, Isabelle</au><au>Proulx, Stéphanie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Culture of human corneal endothelial cells isolated from corneas with Fuchs endothelial corneal dystrophy</atitle><jtitle>Experimental eye research</jtitle><addtitle>Exp Eye Res</addtitle><date>2012</date><risdate>2012</risdate><volume>94</volume><issue>1</issue><spage>22</spage><epage>31</epage><pages>22-31</pages><issn>0014-4835</issn><eissn>1096-0007</eissn><abstract>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.</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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