Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay
The most frequently used parameters to describe the barrier properties of endothelial cells (ECs) in vitro are (i) the macromolecular permeability, indicating the flux of a macromolecular tracer across the endothelium and (ii) electrical impedance of ECs grown on gold-film electrodes reporting on th...
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| Veröffentlicht in: | Scientific reports 2016-03, Vol.6 (1), p.23671-23671, Article 23671 |
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| creator | Bischoff, Iris Hornburger, Michael C. Mayer, Bettina A. Beyerle, Andrea Wegener, Joachim Fürst, Robert |
| description | The most frequently used parameters to describe the barrier properties of endothelial cells (ECs)
in vitro
are (i) the macromolecular permeability, indicating the flux of a macromolecular tracer across the endothelium and (ii) electrical impedance of ECs grown on gold-film electrodes reporting on the cell layer’s tightness for ion flow. Due to the experimental differences between these approaches, inconsistent observations have been described. Here, we present the first direct comparison of these assays applied to one single cell type (human microvascular ECs) under the same experimental conditions. The impact of different pharmacological tools (histamine, forskolin, Y-27632, blebbistatin, TRAP) on endothelial barrier function was analyzed by Transwell
®
tracer assays and two commercial impedance devices (xCELLigence
®
, ECIS
®
). The two impedance techniques provided very similar results for all compounds, whereas macromolecular permeability readings were found to be partly inconsistent with impedance. Possible reasons for these discrepancies are discussed. We conclude that the complementary combination of both approaches is highly recommended to overcome the restrictions of each assay. Since the nature of the growth support may contribute to the observed differences, structure-function relationships should be based on cells that are consistently grown on either permeable or impermeable growth supports in all experiments. |
| doi_str_mv | 10.1038/srep23671 |
| format | Article |
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in vitro
are (i) the macromolecular permeability, indicating the flux of a macromolecular tracer across the endothelium and (ii) electrical impedance of ECs grown on gold-film electrodes reporting on the cell layer’s tightness for ion flow. Due to the experimental differences between these approaches, inconsistent observations have been described. Here, we present the first direct comparison of these assays applied to one single cell type (human microvascular ECs) under the same experimental conditions. The impact of different pharmacological tools (histamine, forskolin, Y-27632, blebbistatin, TRAP) on endothelial barrier function was analyzed by Transwell
®
tracer assays and two commercial impedance devices (xCELLigence
®
, ECIS
®
). The two impedance techniques provided very similar results for all compounds, whereas macromolecular permeability readings were found to be partly inconsistent with impedance. Possible reasons for these discrepancies are discussed. We conclude that the complementary combination of both approaches is highly recommended to overcome the restrictions of each assay. Since the nature of the growth support may contribute to the observed differences, structure-function relationships should be based on cells that are consistently grown on either permeable or impermeable growth supports in all experiments.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/srep23671</identifier><identifier>PMID: 27025965</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/1647/1888 ; 631/57/2283 ; 631/80 ; 692/4019/592 ; Amides - pharmacology ; Biological Assay ; Capillary Permeability - drug effects ; Cells, Cultured ; Electric Impedance ; Electrical impedance ; Endothelial cells ; Endothelial Cells - physiology ; Endothelium ; Endothelium, Vascular - cytology ; Endothelium, Vascular - metabolism ; Forskolin ; Histamine ; Histamine - pharmacology ; Humanities and Social Sciences ; Humans ; Impedance ; Macromolecules ; Microvasculature ; multidisciplinary ; Permeability ; Pyridines - pharmacology ; Science ; Structure-function relationships</subject><ispartof>Scientific reports, 2016-03, Vol.6 (1), p.23671-23671, Article 23671</ispartof><rights>The Author(s) 2016</rights><rights>Copyright Nature Publishing Group Mar 2016</rights><rights>Copyright © 2016, Macmillan Publishers Limited 2016 Macmillan Publishers Limited</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c504t-3596ad224e797005ca54e924749f709f375430d654caf12ca1aa265961a277683</citedby><cites>FETCH-LOGICAL-c504t-3596ad224e797005ca54e924749f709f375430d654caf12ca1aa265961a277683</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4877919/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4877919/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,41120,42189,51576,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27025965$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bischoff, Iris</creatorcontrib><creatorcontrib>Hornburger, Michael C.</creatorcontrib><creatorcontrib>Mayer, Bettina A.</creatorcontrib><creatorcontrib>Beyerle, Andrea</creatorcontrib><creatorcontrib>Wegener, Joachim</creatorcontrib><creatorcontrib>Fürst, Robert</creatorcontrib><title>Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>The most frequently used parameters to describe the barrier properties of endothelial cells (ECs)
in vitro
are (i) the macromolecular permeability, indicating the flux of a macromolecular tracer across the endothelium and (ii) electrical impedance of ECs grown on gold-film electrodes reporting on the cell layer’s tightness for ion flow. Due to the experimental differences between these approaches, inconsistent observations have been described. Here, we present the first direct comparison of these assays applied to one single cell type (human microvascular ECs) under the same experimental conditions. The impact of different pharmacological tools (histamine, forskolin, Y-27632, blebbistatin, TRAP) on endothelial barrier function was analyzed by Transwell
®
tracer assays and two commercial impedance devices (xCELLigence
®
, ECIS
®
). The two impedance techniques provided very similar results for all compounds, whereas macromolecular permeability readings were found to be partly inconsistent with impedance. Possible reasons for these discrepancies are discussed. We conclude that the complementary combination of both approaches is highly recommended to overcome the restrictions of each assay. Since the nature of the growth support may contribute to the observed differences, structure-function relationships should be based on cells that are consistently grown on either permeable or impermeable growth supports in all experiments.</description><subject>631/1647/1888</subject><subject>631/57/2283</subject><subject>631/80</subject><subject>692/4019/592</subject><subject>Amides - pharmacology</subject><subject>Biological Assay</subject><subject>Capillary Permeability - drug effects</subject><subject>Cells, Cultured</subject><subject>Electric Impedance</subject><subject>Electrical impedance</subject><subject>Endothelial cells</subject><subject>Endothelial Cells - physiology</subject><subject>Endothelium</subject><subject>Endothelium, Vascular - cytology</subject><subject>Endothelium, Vascular - metabolism</subject><subject>Forskolin</subject><subject>Histamine</subject><subject>Histamine - pharmacology</subject><subject>Humanities and Social Sciences</subject><subject>Humans</subject><subject>Impedance</subject><subject>Macromolecules</subject><subject>Microvasculature</subject><subject>multidisciplinary</subject><subject>Permeability</subject><subject>Pyridines - pharmacology</subject><subject>Science</subject><subject>Structure-function relationships</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNplkcFu1DAQhiMEolXpgRdAlrhApZTYceKYAxKqKFSqBAc4W7POZOvKcRZPslLfgMdm6JbVQn2xpfnnm3_8F8VLWZ3Lqu7eUcaNqlsjnxTHqtJNqWqlnh68j4pTotuKT6OslvZ5caRMpRrbNsfFr29hHiBGEiEJIEKikNZiDD5PWyC_RMgCUz_NNxgDRLGCnANmMSzJz2FK70UYN9hD8liugLAXPW6DRxJbzLSQ4EbhI6ODFyMwdpwi7rhzBs8orsHdi-IZ-yA8fbhPih-Xn75ffCmvv36-uvh4Xfqm0nNZs23oldJorOGNPDQardJG28FUdqhNo-uqbxvtYZDKgwRQLTdJUMa0XX1SfNhxN8tqxN5jYhfRbXIYId-5CYL7t5LCjVtPW6c7Y6y0DHjzAMjTzwVpdmMgjzFCwmkhJ40xumtsJ1n6-j_p7bTkxOs52dmu7Yy6d_R2p-K_IU5z2JuRlfsTsdtHzNpXh-73yr-BsuBsJyAupTXmg5GPaL8Bvvmy-w</recordid><startdate>20160330</startdate><enddate>20160330</enddate><creator>Bischoff, Iris</creator><creator>Hornburger, Michael C.</creator><creator>Mayer, Bettina A.</creator><creator>Beyerle, Andrea</creator><creator>Wegener, Joachim</creator><creator>Fürst, Robert</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</scope><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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20160330</creationdate><title>Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay</title><author>Bischoff, Iris ; Hornburger, Michael C. ; Mayer, Bettina A. ; Beyerle, Andrea ; Wegener, Joachim ; Fürst, Robert</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c504t-3596ad224e797005ca54e924749f709f375430d654caf12ca1aa265961a277683</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>631/1647/1888</topic><topic>631/57/2283</topic><topic>631/80</topic><topic>692/4019/592</topic><topic>Amides - pharmacology</topic><topic>Biological Assay</topic><topic>Capillary Permeability - drug effects</topic><topic>Cells, Cultured</topic><topic>Electric Impedance</topic><topic>Electrical impedance</topic><topic>Endothelial cells</topic><topic>Endothelial Cells - physiology</topic><topic>Endothelium</topic><topic>Endothelium, Vascular - cytology</topic><topic>Endothelium, Vascular - metabolism</topic><topic>Forskolin</topic><topic>Histamine</topic><topic>Histamine - pharmacology</topic><topic>Humanities and Social Sciences</topic><topic>Humans</topic><topic>Impedance</topic><topic>Macromolecules</topic><topic>Microvasculature</topic><topic>multidisciplinary</topic><topic>Permeability</topic><topic>Pyridines - pharmacology</topic><topic>Science</topic><topic>Structure-function relationships</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bischoff, Iris</creatorcontrib><creatorcontrib>Hornburger, Michael C.</creatorcontrib><creatorcontrib>Mayer, Bettina A.</creatorcontrib><creatorcontrib>Beyerle, Andrea</creatorcontrib><creatorcontrib>Wegener, Joachim</creatorcontrib><creatorcontrib>Fürst, Robert</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bischoff, Iris</au><au>Hornburger, Michael C.</au><au>Mayer, Bettina A.</au><au>Beyerle, Andrea</au><au>Wegener, Joachim</au><au>Fürst, Robert</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2016-03-30</date><risdate>2016</risdate><volume>6</volume><issue>1</issue><spage>23671</spage><epage>23671</epage><pages>23671-23671</pages><artnum>23671</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>The most frequently used parameters to describe the barrier properties of endothelial cells (ECs)
in vitro
are (i) the macromolecular permeability, indicating the flux of a macromolecular tracer across the endothelium and (ii) electrical impedance of ECs grown on gold-film electrodes reporting on the cell layer’s tightness for ion flow. Due to the experimental differences between these approaches, inconsistent observations have been described. Here, we present the first direct comparison of these assays applied to one single cell type (human microvascular ECs) under the same experimental conditions. The impact of different pharmacological tools (histamine, forskolin, Y-27632, blebbistatin, TRAP) on endothelial barrier function was analyzed by Transwell
®
tracer assays and two commercial impedance devices (xCELLigence
®
, ECIS
®
). The two impedance techniques provided very similar results for all compounds, whereas macromolecular permeability readings were found to be partly inconsistent with impedance. Possible reasons for these discrepancies are discussed. We conclude that the complementary combination of both approaches is highly recommended to overcome the restrictions of each assay. Since the nature of the growth support may contribute to the observed differences, structure-function relationships should be based on cells that are consistently grown on either permeable or impermeable growth supports in all experiments.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>27025965</pmid><doi>10.1038/srep23671</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 631/1647/1888 631/57/2283 631/80 692/4019/592 Amides - pharmacology Biological Assay Capillary Permeability - drug effects Cells, Cultured Electric Impedance Electrical impedance Endothelial cells Endothelial Cells - physiology Endothelium Endothelium, Vascular - cytology Endothelium, Vascular - metabolism Forskolin Histamine Histamine - pharmacology Humanities and Social Sciences Humans Impedance Macromolecules Microvasculature multidisciplinary Permeability Pyridines - pharmacology Science Structure-function relationships |
| title | Pitfalls in assessing microvascular endothelial barrier function: impedance-based devices versus the classic macromolecular tracer assay |
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