Development of immortalized rat conjunctival epithelial cell lines: An in vitro model to examine transepithelial antigen delivery
The objective of these studies was to develop conjunctival epithelial cell lines for investigation of antigen translocation across a mucosal barrier. Conjunctival epithelial cells from Fischer 344 rats were immortalized with pSV3(neo) resulting in two cell lines—CJ4.1A and CJ4.3C. Each formed conflu...
Gespeichert in:
Veröffentlicht in: | Experimental eye research 2007-02, Vol.84 (2), p.323-331 |
---|---|
Hauptverfasser: | , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 331 |
---|---|
container_issue | 2 |
container_start_page | 323 |
container_title | Experimental eye research |
container_volume | 84 |
creator | O'Sullivan, Nancy L. Baylor, Alfred E. Montgomery, Paul C. |
description | The objective of these studies was to develop conjunctival epithelial cell lines for investigation of antigen translocation across a mucosal barrier. Conjunctival epithelial cells from Fischer 344 rats were immortalized with pSV3(neo) resulting in two cell lines—CJ4.1A and CJ4.3C. Each formed confluent cell layers with epithelial morphology when grown on permeable membrane filters. They expressed the SV40 T antigen, the conjunctiva-specific cytokeratin 4, the goblet cell-specific cytokeratin 7 and were negative for the corneal epithelial cell-specific cytokeratin 12. The cell lines have been in culture for over 60 passages, and the population doubling times were 22
±
7
h for CJ4.1A and 23
±
9
h for CJ4.3C. When grown on Transwell™ membranes, each cell line achieved a transepithelial electrical resistance of 600–800 Ω
cm
2 by 3–4
days and maintained a high resistance for several days. Both cell lines expressed zona occludens-1 at confluence. At 24
h following addition of 250
μg of FITC-labeled ovalbumin to the apical chambers, 15
±
6
μg could be detected in the basal chamber of CJ4.1A and 6
±
1
μg in the basal medium of CJ4.3C. In contrast, 82
±
6
μg was detected in the lower chambers of cell-free Transwells. Similarly, Transwells containing confluent CJ4.1A or CJ4.3C cells impeded passage of 0.1
μm diameter polystyrene microspheres (5
±
1% and 4
±
1%, respectively, of the apical input), compared to 26
±
6% of the input microspheres recovered from the basal chambers of cell-free Transwells. Pretreatment with 4
mM EGTA for 10
min caused an increase in OVA-FITC translocation across CJ4.3C cells. Incubation in the presence of 4
mM EGTA significantly increased OVA-FITC translocation across both cell lines, relative to untreated cell layers. Morphological and functional characterization indicates that these cells provide a useful experimental tool to assess strategies for enhancing transepithelial antigen uptake. |
doi_str_mv | 10.1016/j.exer.2006.10.005 |
format | Article |
fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1839946</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0014483506003988</els_id><sourcerecordid>68393110</sourcerecordid><originalsourceid>FETCH-LOGICAL-c453t-fc12bbd15658e835a2cf96d58b8abd582306fd153a8a4a73a968f4f8c2241adf3</originalsourceid><addsrcrecordid>eNp9UUtv1DAQthAVXQp_gAPyiVsWO3a8CUJIVctLqtRLe7YcZ9LOyrEX2xu13PjnONoVlAunseZ7jGc-Qt5wtuaMq_fbNTxAXNeMqdJYM9Y8IyvOOlUxxjbPyYoxLivZiuaUvExpW7pCbuQLcso3vBYNVyvy6xJmcGE3gc80jBSnKcRsHP6EgUaTqQ1-u_c242wchR3me3BYnhacow49pA_03FP0dMYcA53CAI7mQOHBTAWmORqfngiNz3gHnhYazhAfX5GT0bgEr4_1jNx--Xxz8a26uv76_eL8qrKyEbkaLa_7fuCNalooK5najp0amrZvTV9KLZgaCyxMa6TZCNOpdpRja-tacjOM4ox8Ovju9v0Egy0LR-P0LuJk4qMOBvW_iMd7fRdmzVvRdVIVg3dHgxh-7CFlPWFazmA8hH3SqvAE56wQ6wPRxpBShPHPEM70kpze6iU5vSS39EpyRfT26ff-So5RFcLHAwHKkWYs8mQRvIUBI9ish4D_8_8NJyKvIQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>68393110</pqid></control><display><type>article</type><title>Development of immortalized rat conjunctival epithelial cell lines: An in vitro model to examine transepithelial antigen delivery</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>O'Sullivan, Nancy L. ; Baylor, Alfred E. ; Montgomery, Paul C.</creator><creatorcontrib>O'Sullivan, Nancy L. ; Baylor, Alfred E. ; Montgomery, Paul C.</creatorcontrib><description>The objective of these studies was to develop conjunctival epithelial cell lines for investigation of antigen translocation across a mucosal barrier. Conjunctival epithelial cells from Fischer 344 rats were immortalized with pSV3(neo) resulting in two cell lines—CJ4.1A and CJ4.3C. Each formed confluent cell layers with epithelial morphology when grown on permeable membrane filters. They expressed the SV40 T antigen, the conjunctiva-specific cytokeratin 4, the goblet cell-specific cytokeratin 7 and were negative for the corneal epithelial cell-specific cytokeratin 12. The cell lines have been in culture for over 60 passages, and the population doubling times were 22
±
7
h for CJ4.1A and 23
±
9
h for CJ4.3C. When grown on Transwell™ membranes, each cell line achieved a transepithelial electrical resistance of 600–800 Ω
cm
2 by 3–4
days and maintained a high resistance for several days. Both cell lines expressed zona occludens-1 at confluence. At 24
h following addition of 250
μg of FITC-labeled ovalbumin to the apical chambers, 15
±
6
μg could be detected in the basal chamber of CJ4.1A and 6
±
1
μg in the basal medium of CJ4.3C. In contrast, 82
±
6
μg was detected in the lower chambers of cell-free Transwells. Similarly, Transwells containing confluent CJ4.1A or CJ4.3C cells impeded passage of 0.1
μm diameter polystyrene microspheres (5
±
1% and 4
±
1%, respectively, of the apical input), compared to 26
±
6% of the input microspheres recovered from the basal chambers of cell-free Transwells. Pretreatment with 4
mM EGTA for 10
min caused an increase in OVA-FITC translocation across CJ4.3C cells. Incubation in the presence of 4
mM EGTA significantly increased OVA-FITC translocation across both cell lines, relative to untreated cell layers. Morphological and functional characterization indicates that these cells provide a useful experimental tool to assess strategies for enhancing transepithelial antigen uptake.</description><identifier>ISSN: 0014-4835</identifier><identifier>EISSN: 1096-0007</identifier><identifier>DOI: 10.1016/j.exer.2006.10.005</identifier><identifier>PMID: 17123516</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Animals ; Antigens - metabolism ; cell culture ; Cell Differentiation ; Cell Line, Transformed ; conjunctiva ; Conjunctiva - cytology ; Conjunctiva - immunology ; Conjunctiva - metabolism ; Epithelial Cells - cytology ; Epithelial Cells - immunology ; Epithelial Cells - metabolism ; Eye Proteins - metabolism ; Keratins - metabolism ; Microscopy, Confocal ; Microscopy, Phase-Contrast ; Microspheres ; Ovalbumin - pharmacokinetics ; rat ; Rats ; Rats, Inbred F344 ; Tight Junctions - metabolism</subject><ispartof>Experimental eye research, 2007-02, Vol.84 (2), p.323-331</ispartof><rights>2006 Elsevier Ltd</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c453t-fc12bbd15658e835a2cf96d58b8abd582306fd153a8a4a73a968f4f8c2241adf3</citedby><cites>FETCH-LOGICAL-c453t-fc12bbd15658e835a2cf96d58b8abd582306fd153a8a4a73a968f4f8c2241adf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.exer.2006.10.005$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17123516$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>O'Sullivan, Nancy L.</creatorcontrib><creatorcontrib>Baylor, Alfred E.</creatorcontrib><creatorcontrib>Montgomery, Paul C.</creatorcontrib><title>Development of immortalized rat conjunctival epithelial cell lines: An in vitro model to examine transepithelial antigen delivery</title><title>Experimental eye research</title><addtitle>Exp Eye Res</addtitle><description>The objective of these studies was to develop conjunctival epithelial cell lines for investigation of antigen translocation across a mucosal barrier. Conjunctival epithelial cells from Fischer 344 rats were immortalized with pSV3(neo) resulting in two cell lines—CJ4.1A and CJ4.3C. Each formed confluent cell layers with epithelial morphology when grown on permeable membrane filters. They expressed the SV40 T antigen, the conjunctiva-specific cytokeratin 4, the goblet cell-specific cytokeratin 7 and were negative for the corneal epithelial cell-specific cytokeratin 12. The cell lines have been in culture for over 60 passages, and the population doubling times were 22
±
7
h for CJ4.1A and 23
±
9
h for CJ4.3C. When grown on Transwell™ membranes, each cell line achieved a transepithelial electrical resistance of 600–800 Ω
cm
2 by 3–4
days and maintained a high resistance for several days. Both cell lines expressed zona occludens-1 at confluence. At 24
h following addition of 250
μg of FITC-labeled ovalbumin to the apical chambers, 15
±
6
μg could be detected in the basal chamber of CJ4.1A and 6
±
1
μg in the basal medium of CJ4.3C. In contrast, 82
±
6
μg was detected in the lower chambers of cell-free Transwells. Similarly, Transwells containing confluent CJ4.1A or CJ4.3C cells impeded passage of 0.1
μm diameter polystyrene microspheres (5
±
1% and 4
±
1%, respectively, of the apical input), compared to 26
±
6% of the input microspheres recovered from the basal chambers of cell-free Transwells. Pretreatment with 4
mM EGTA for 10
min caused an increase in OVA-FITC translocation across CJ4.3C cells. Incubation in the presence of 4
mM EGTA significantly increased OVA-FITC translocation across both cell lines, relative to untreated cell layers. Morphological and functional characterization indicates that these cells provide a useful experimental tool to assess strategies for enhancing transepithelial antigen uptake.</description><subject>Animals</subject><subject>Antigens - metabolism</subject><subject>cell culture</subject><subject>Cell Differentiation</subject><subject>Cell Line, Transformed</subject><subject>conjunctiva</subject><subject>Conjunctiva - cytology</subject><subject>Conjunctiva - immunology</subject><subject>Conjunctiva - metabolism</subject><subject>Epithelial Cells - cytology</subject><subject>Epithelial Cells - immunology</subject><subject>Epithelial Cells - metabolism</subject><subject>Eye Proteins - metabolism</subject><subject>Keratins - metabolism</subject><subject>Microscopy, Confocal</subject><subject>Microscopy, Phase-Contrast</subject><subject>Microspheres</subject><subject>Ovalbumin - pharmacokinetics</subject><subject>rat</subject><subject>Rats</subject><subject>Rats, Inbred F344</subject><subject>Tight Junctions - metabolism</subject><issn>0014-4835</issn><issn>1096-0007</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9UUtv1DAQthAVXQp_gAPyiVsWO3a8CUJIVctLqtRLe7YcZ9LOyrEX2xu13PjnONoVlAunseZ7jGc-Qt5wtuaMq_fbNTxAXNeMqdJYM9Y8IyvOOlUxxjbPyYoxLivZiuaUvExpW7pCbuQLcso3vBYNVyvy6xJmcGE3gc80jBSnKcRsHP6EgUaTqQ1-u_c242wchR3me3BYnhacow49pA_03FP0dMYcA53CAI7mQOHBTAWmORqfngiNz3gHnhYazhAfX5GT0bgEr4_1jNx--Xxz8a26uv76_eL8qrKyEbkaLa_7fuCNalooK5najp0amrZvTV9KLZgaCyxMa6TZCNOpdpRja-tacjOM4ox8Ovju9v0Egy0LR-P0LuJk4qMOBvW_iMd7fRdmzVvRdVIVg3dHgxh-7CFlPWFazmA8hH3SqvAE56wQ6wPRxpBShPHPEM70kpze6iU5vSS39EpyRfT26ff-So5RFcLHAwHKkWYs8mQRvIUBI9ish4D_8_8NJyKvIQ</recordid><startdate>20070201</startdate><enddate>20070201</enddate><creator>O'Sullivan, Nancy L.</creator><creator>Baylor, Alfred E.</creator><creator>Montgomery, Paul C.</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><scope>5PM</scope></search><sort><creationdate>20070201</creationdate><title>Development of immortalized rat conjunctival epithelial cell lines: An in vitro model to examine transepithelial antigen delivery</title><author>O'Sullivan, Nancy L. ; Baylor, Alfred E. ; Montgomery, Paul C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c453t-fc12bbd15658e835a2cf96d58b8abd582306fd153a8a4a73a968f4f8c2241adf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Antigens - metabolism</topic><topic>cell culture</topic><topic>Cell Differentiation</topic><topic>Cell Line, Transformed</topic><topic>conjunctiva</topic><topic>Conjunctiva - cytology</topic><topic>Conjunctiva - immunology</topic><topic>Conjunctiva - metabolism</topic><topic>Epithelial Cells - cytology</topic><topic>Epithelial Cells - immunology</topic><topic>Epithelial Cells - metabolism</topic><topic>Eye Proteins - metabolism</topic><topic>Keratins - metabolism</topic><topic>Microscopy, Confocal</topic><topic>Microscopy, Phase-Contrast</topic><topic>Microspheres</topic><topic>Ovalbumin - pharmacokinetics</topic><topic>rat</topic><topic>Rats</topic><topic>Rats, Inbred F344</topic><topic>Tight Junctions - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>O'Sullivan, Nancy L.</creatorcontrib><creatorcontrib>Baylor, Alfred E.</creatorcontrib><creatorcontrib>Montgomery, Paul C.</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><collection>PubMed Central (Full Participant titles)</collection><jtitle>Experimental eye research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>O'Sullivan, Nancy L.</au><au>Baylor, Alfred E.</au><au>Montgomery, Paul C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of immortalized rat conjunctival epithelial cell lines: An in vitro model to examine transepithelial antigen delivery</atitle><jtitle>Experimental eye research</jtitle><addtitle>Exp Eye Res</addtitle><date>2007-02-01</date><risdate>2007</risdate><volume>84</volume><issue>2</issue><spage>323</spage><epage>331</epage><pages>323-331</pages><issn>0014-4835</issn><eissn>1096-0007</eissn><abstract>The objective of these studies was to develop conjunctival epithelial cell lines for investigation of antigen translocation across a mucosal barrier. Conjunctival epithelial cells from Fischer 344 rats were immortalized with pSV3(neo) resulting in two cell lines—CJ4.1A and CJ4.3C. Each formed confluent cell layers with epithelial morphology when grown on permeable membrane filters. They expressed the SV40 T antigen, the conjunctiva-specific cytokeratin 4, the goblet cell-specific cytokeratin 7 and were negative for the corneal epithelial cell-specific cytokeratin 12. The cell lines have been in culture for over 60 passages, and the population doubling times were 22
±
7
h for CJ4.1A and 23
±
9
h for CJ4.3C. When grown on Transwell™ membranes, each cell line achieved a transepithelial electrical resistance of 600–800 Ω
cm
2 by 3–4
days and maintained a high resistance for several days. Both cell lines expressed zona occludens-1 at confluence. At 24
h following addition of 250
μg of FITC-labeled ovalbumin to the apical chambers, 15
±
6
μg could be detected in the basal chamber of CJ4.1A and 6
±
1
μg in the basal medium of CJ4.3C. In contrast, 82
±
6
μg was detected in the lower chambers of cell-free Transwells. Similarly, Transwells containing confluent CJ4.1A or CJ4.3C cells impeded passage of 0.1
μm diameter polystyrene microspheres (5
±
1% and 4
±
1%, respectively, of the apical input), compared to 26
±
6% of the input microspheres recovered from the basal chambers of cell-free Transwells. Pretreatment with 4
mM EGTA for 10
min caused an increase in OVA-FITC translocation across CJ4.3C cells. Incubation in the presence of 4
mM EGTA significantly increased OVA-FITC translocation across both cell lines, relative to untreated cell layers. Morphological and functional characterization indicates that these cells provide a useful experimental tool to assess strategies for enhancing transepithelial antigen uptake.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>17123516</pmid><doi>10.1016/j.exer.2006.10.005</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0014-4835 |
ispartof | Experimental eye research, 2007-02, Vol.84 (2), p.323-331 |
issn | 0014-4835 1096-0007 |
language | eng |
recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1839946 |
source | MEDLINE; Access via ScienceDirect (Elsevier) |
subjects | Animals Antigens - metabolism cell culture Cell Differentiation Cell Line, Transformed conjunctiva Conjunctiva - cytology Conjunctiva - immunology Conjunctiva - metabolism Epithelial Cells - cytology Epithelial Cells - immunology Epithelial Cells - metabolism Eye Proteins - metabolism Keratins - metabolism Microscopy, Confocal Microscopy, Phase-Contrast Microspheres Ovalbumin - pharmacokinetics rat Rats Rats, Inbred F344 Tight Junctions - metabolism |
title | Development of immortalized rat conjunctival epithelial cell lines: An in vitro model to examine transepithelial antigen delivery |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T17%3A01%3A50IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20of%20immortalized%20rat%20conjunctival%20epithelial%20cell%20lines:%20An%20in%20vitro%20model%20to%20examine%20transepithelial%20antigen%20delivery&rft.jtitle=Experimental%20eye%20research&rft.au=O'Sullivan,%20Nancy%20L.&rft.date=2007-02-01&rft.volume=84&rft.issue=2&rft.spage=323&rft.epage=331&rft.pages=323-331&rft.issn=0014-4835&rft.eissn=1096-0007&rft_id=info:doi/10.1016/j.exer.2006.10.005&rft_dat=%3Cproquest_pubme%3E68393110%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=68393110&rft_id=info:pmid/17123516&rft_els_id=S0014483506003988&rfr_iscdi=true |