Detection of Epithelial-Cell Injury, and Quantification of Infection, in the HCT-8 Organoid Model of Cryptosporidiosis
Background. Intestinal cells grown in microgravity produce a three-dimensional tissue assembly, or “organoid,” similar to the human intestinal mucosa, making it an ideal model for enteric infections such as cryptosporidiosis. Methods. HCT-8 cells were grown in a reduced-gravity, low-shear, rotating-...
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| Veröffentlicht in: | The Journal of Infectious Diseases 2008-07, Vol.198 (1), p.143-149 |
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| creator | Alcantara Warren, Cirle Destura, Raul V. Sevilleja, Jesus Emmanuel A. D. Barroso, Luis F. Carvalho, Humberto Barrett, Leah J. O'Brien, Alison D. Guerrant, Richard L. |
| description | Background. Intestinal cells grown in microgravity produce a three-dimensional tissue assembly, or “organoid,” similar to the human intestinal mucosa, making it an ideal model for enteric infections such as cryptosporidiosis. Methods. HCT-8 cells were grown in a reduced-gravity, low-shear, rotating-wall vessel (RWV) and were infected with Cryptosporidium parvum oocysts. Routine and electron microscopy (EM), immunolabeling with fluoresceinlabeled Vicia villosa lectin and phycoerythrin-labeled monoclonal antibody to a 15-kD surface-membrane protein, and quantitative polymerase chain reaction (qPCR) using probes for 18s rRNA of C. parvum and HCT-8 cells were performed. Results. The RWV allowed development of columnar epithelium-like structures. Higher magnification revealed well-developed brush borders at the apical side of the tissue. Incubation with C. parvum resulted in patchy disruption of the epithelium and, at the surface of several epithelial cells, in localized infection with the organism. EM revealed irregular stunting of microvilli, foci of indistinct tight junctions, and areas of loose paracellular spaces. qPCR showed a 1.85-log (i.e., 70-fold) progression of infection from 6 h to 48 h of incubation. Conclusion. The HCT-8 organoid displayed morphologic changes indicative of successful and quantifiable infection with C. parvum. The HCT-8 organoid-culture system may have application in interventional in vitro studies of cryptosporidiosis. |
| doi_str_mv | 10.1086/588819 |
| format | Article |
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D. ; Barroso, Luis F. ; Carvalho, Humberto ; Barrett, Leah J. ; O'Brien, Alison D. ; Guerrant, Richard L.</creator><creatorcontrib>Alcantara Warren, Cirle ; Destura, Raul V. ; Sevilleja, Jesus Emmanuel A. D. ; Barroso, Luis F. ; Carvalho, Humberto ; Barrett, Leah J. ; O'Brien, Alison D. ; Guerrant, Richard L.</creatorcontrib><description>Background. Intestinal cells grown in microgravity produce a three-dimensional tissue assembly, or “organoid,” similar to the human intestinal mucosa, making it an ideal model for enteric infections such as cryptosporidiosis. Methods. HCT-8 cells were grown in a reduced-gravity, low-shear, rotating-wall vessel (RWV) and were infected with Cryptosporidium parvum oocysts. Routine and electron microscopy (EM), immunolabeling with fluoresceinlabeled Vicia villosa lectin and phycoerythrin-labeled monoclonal antibody to a 15-kD surface-membrane protein, and quantitative polymerase chain reaction (qPCR) using probes for 18s rRNA of C. parvum and HCT-8 cells were performed. Results. The RWV allowed development of columnar epithelium-like structures. Higher magnification revealed well-developed brush borders at the apical side of the tissue. Incubation with C. parvum resulted in patchy disruption of the epithelium and, at the surface of several epithelial cells, in localized infection with the organism. EM revealed irregular stunting of microvilli, foci of indistinct tight junctions, and areas of loose paracellular spaces. qPCR showed a 1.85-log (i.e., 70-fold) progression of infection from 6 h to 48 h of incubation. Conclusion. The HCT-8 organoid displayed morphologic changes indicative of successful and quantifiable infection with C. parvum. The HCT-8 organoid-culture system may have application in interventional in vitro studies of cryptosporidiosis.</description><identifier>ISSN: 0022-1899</identifier><identifier>EISSN: 1537-6591</identifier><identifier>EISSN: 1537-6613</identifier><identifier>DOI: 10.1086/588819</identifier><identifier>PMID: 18498239</identifier><identifier>CODEN: JIDIAQ</identifier><language>eng</language><publisher>Chicago, IL: The University Chicago Press</publisher><subject>Animals ; Biological and medical sciences ; Cell Culture Techniques ; Cell Line, Tumor ; Cell lines ; Cryptosporidiosis ; Cryptosporidiosis - parasitology ; Cryptosporidiosis - pathology ; Cryptosporidium parvum ; Cryptosporidium parvum - physiology ; Epithelial cells ; Epithelial Cells - parasitology ; Epithelial Cells - pathology ; Fundamental and applied biological sciences. Psychology ; Human protozoal diseases ; Humans ; Infections ; Infectious diseases ; Medical sciences ; Microbiology ; Microgravity ; Microvilli ; Oocysts ; Organoids ; Organoids - cytology ; Organoids - parasitology ; Parasites ; Parasitic diseases ; Polymerase chain reaction ; Protozoal diseases ; Rotation ; Vicia villosa</subject><ispartof>The Journal of Infectious Diseases, 2008-07, Vol.198 (1), p.143-149</ispartof><rights>Copyright 2008 Infectious Diseases Society of America</rights><rights>2008 by the Infectious Diseases Society of America. 2008</rights><rights>2008 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c487t-69f6c7adca0ef107fc4f9b1b16016704945b7a74c21d5f96f1bc85673ba05c103</citedby><cites>FETCH-LOGICAL-c487t-69f6c7adca0ef107fc4f9b1b16016704945b7a74c21d5f96f1bc85673ba05c103</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/40254005$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/40254005$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>314,780,784,803,27922,27923,58015,58248</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20427175$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18498239$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Alcantara Warren, Cirle</creatorcontrib><creatorcontrib>Destura, Raul V.</creatorcontrib><creatorcontrib>Sevilleja, Jesus Emmanuel A. D.</creatorcontrib><creatorcontrib>Barroso, Luis F.</creatorcontrib><creatorcontrib>Carvalho, Humberto</creatorcontrib><creatorcontrib>Barrett, Leah J.</creatorcontrib><creatorcontrib>O'Brien, Alison D.</creatorcontrib><creatorcontrib>Guerrant, Richard L.</creatorcontrib><title>Detection of Epithelial-Cell Injury, and Quantification of Infection, in the HCT-8 Organoid Model of Cryptosporidiosis</title><title>The Journal of Infectious Diseases</title><addtitle>The Journal of Infectious Diseases</addtitle><addtitle>The Journal of Infectious Diseases</addtitle><description>Background. Intestinal cells grown in microgravity produce a three-dimensional tissue assembly, or “organoid,” similar to the human intestinal mucosa, making it an ideal model for enteric infections such as cryptosporidiosis. Methods. HCT-8 cells were grown in a reduced-gravity, low-shear, rotating-wall vessel (RWV) and were infected with Cryptosporidium parvum oocysts. Routine and electron microscopy (EM), immunolabeling with fluoresceinlabeled Vicia villosa lectin and phycoerythrin-labeled monoclonal antibody to a 15-kD surface-membrane protein, and quantitative polymerase chain reaction (qPCR) using probes for 18s rRNA of C. parvum and HCT-8 cells were performed. Results. The RWV allowed development of columnar epithelium-like structures. Higher magnification revealed well-developed brush borders at the apical side of the tissue. Incubation with C. parvum resulted in patchy disruption of the epithelium and, at the surface of several epithelial cells, in localized infection with the organism. EM revealed irregular stunting of microvilli, foci of indistinct tight junctions, and areas of loose paracellular spaces. qPCR showed a 1.85-log (i.e., 70-fold) progression of infection from 6 h to 48 h of incubation. Conclusion. The HCT-8 organoid displayed morphologic changes indicative of successful and quantifiable infection with C. parvum. The HCT-8 organoid-culture system may have application in interventional in vitro studies of cryptosporidiosis.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell Culture Techniques</subject><subject>Cell Line, Tumor</subject><subject>Cell lines</subject><subject>Cryptosporidiosis</subject><subject>Cryptosporidiosis - parasitology</subject><subject>Cryptosporidiosis - pathology</subject><subject>Cryptosporidium parvum</subject><subject>Cryptosporidium parvum - physiology</subject><subject>Epithelial cells</subject><subject>Epithelial Cells - parasitology</subject><subject>Epithelial Cells - pathology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Human protozoal diseases</subject><subject>Humans</subject><subject>Infections</subject><subject>Infectious diseases</subject><subject>Medical sciences</subject><subject>Microbiology</subject><subject>Microgravity</subject><subject>Microvilli</subject><subject>Oocysts</subject><subject>Organoids</subject><subject>Organoids - cytology</subject><subject>Organoids - parasitology</subject><subject>Parasites</subject><subject>Parasitic diseases</subject><subject>Polymerase chain reaction</subject><subject>Protozoal diseases</subject><subject>Rotation</subject><subject>Vicia villosa</subject><issn>0022-1899</issn><issn>1537-6591</issn><issn>1537-6613</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0U1v1DAQBuAIgehS4B-AzAFODYwTfx5RaHcXFapKBaFeLMexwUs2DnaC2H9PVlm2JwS--DDPjOV5s-wphtcYBHtDhRBY3ssWmJY8Z1Ti-9kCoChyLKQ8yR6ltAHAWAB9mJ1gQaQoSrnIfr6zgzWDDx0KDp33fvhmW6_bvLJti9bdZoy7M6S7Bl2Puhu880b_0evOza1nyHdoakSr6iYX6Cp-1V3wDfoQGtvuZRV3_RBSH6JvfEg-Pc4eON0m--Rwn2afLs5vqlV-ebVcV28vc0MEH3ImHTNcN0aDdRi4M8TJGteYAWYciCS05poTU-CGOskcro2gjJe1BmowlKfZq3luH8OP0aZBbX0y09d0Z8OYFMdsOtMq_gWx5JQUDP4HEg6U30ETQ0rROtVHv9VxpzCofWZqzmyCzw8Tx3prmzt2CGkCLw9AJ6NbF3VnfDq6AkjBMaeTezG7MPZ_f-zZbDZpCPGoCBSUAOxn5HPdp8H-OtZ1_K6mtXKqVl9u1fKCvb--XX5Un8vfX92_7A</recordid><startdate>20080701</startdate><enddate>20080701</enddate><creator>Alcantara Warren, Cirle</creator><creator>Destura, Raul V.</creator><creator>Sevilleja, Jesus Emmanuel A. D.</creator><creator>Barroso, Luis F.</creator><creator>Carvalho, Humberto</creator><creator>Barrett, Leah J.</creator><creator>O'Brien, Alison D.</creator><creator>Guerrant, Richard L.</creator><general>The University Chicago Press</general><general>University of Chicago Press</general><scope>BSCLL</scope><scope>IQODW</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>7UA</scope><scope>C1K</scope><scope>M7N</scope><scope>F1W</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope><scope>7X8</scope></search><sort><creationdate>20080701</creationdate><title>Detection of Epithelial-Cell Injury, and Quantification of Infection, in the HCT-8 Organoid Model of Cryptosporidiosis</title><author>Alcantara Warren, Cirle ; Destura, Raul V. ; Sevilleja, Jesus Emmanuel A. 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Psychology</topic><topic>Human protozoal diseases</topic><topic>Humans</topic><topic>Infections</topic><topic>Infectious diseases</topic><topic>Medical sciences</topic><topic>Microbiology</topic><topic>Microgravity</topic><topic>Microvilli</topic><topic>Oocysts</topic><topic>Organoids</topic><topic>Organoids - cytology</topic><topic>Organoids - parasitology</topic><topic>Parasites</topic><topic>Parasitic diseases</topic><topic>Polymerase chain reaction</topic><topic>Protozoal diseases</topic><topic>Rotation</topic><topic>Vicia villosa</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Alcantara Warren, Cirle</creatorcontrib><creatorcontrib>Destura, Raul V.</creatorcontrib><creatorcontrib>Sevilleja, Jesus Emmanuel A. D.</creatorcontrib><creatorcontrib>Barroso, Luis F.</creatorcontrib><creatorcontrib>Carvalho, Humberto</creatorcontrib><creatorcontrib>Barrett, Leah J.</creatorcontrib><creatorcontrib>O'Brien, Alison D.</creatorcontrib><creatorcontrib>Guerrant, Richard L.</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Water Resources Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of Infectious Diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Alcantara Warren, Cirle</au><au>Destura, Raul V.</au><au>Sevilleja, Jesus Emmanuel A. D.</au><au>Barroso, Luis F.</au><au>Carvalho, Humberto</au><au>Barrett, Leah J.</au><au>O'Brien, Alison D.</au><au>Guerrant, Richard L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of Epithelial-Cell Injury, and Quantification of Infection, in the HCT-8 Organoid Model of Cryptosporidiosis</atitle><jtitle>The Journal of Infectious Diseases</jtitle><stitle>The Journal of Infectious Diseases</stitle><addtitle>The Journal of Infectious Diseases</addtitle><date>2008-07-01</date><risdate>2008</risdate><volume>198</volume><issue>1</issue><spage>143</spage><epage>149</epage><pages>143-149</pages><issn>0022-1899</issn><eissn>1537-6591</eissn><eissn>1537-6613</eissn><coden>JIDIAQ</coden><abstract>Background. Intestinal cells grown in microgravity produce a three-dimensional tissue assembly, or “organoid,” similar to the human intestinal mucosa, making it an ideal model for enteric infections such as cryptosporidiosis. Methods. HCT-8 cells were grown in a reduced-gravity, low-shear, rotating-wall vessel (RWV) and were infected with Cryptosporidium parvum oocysts. Routine and electron microscopy (EM), immunolabeling with fluoresceinlabeled Vicia villosa lectin and phycoerythrin-labeled monoclonal antibody to a 15-kD surface-membrane protein, and quantitative polymerase chain reaction (qPCR) using probes for 18s rRNA of C. parvum and HCT-8 cells were performed. Results. The RWV allowed development of columnar epithelium-like structures. Higher magnification revealed well-developed brush borders at the apical side of the tissue. Incubation with C. parvum resulted in patchy disruption of the epithelium and, at the surface of several epithelial cells, in localized infection with the organism. EM revealed irregular stunting of microvilli, foci of indistinct tight junctions, and areas of loose paracellular spaces. qPCR showed a 1.85-log (i.e., 70-fold) progression of infection from 6 h to 48 h of incubation. Conclusion. The HCT-8 organoid displayed morphologic changes indicative of successful and quantifiable infection with C. parvum. The HCT-8 organoid-culture system may have application in interventional in vitro studies of cryptosporidiosis.</abstract><cop>Chicago, IL</cop><pub>The University Chicago Press</pub><pmid>18498239</pmid><doi>10.1086/588819</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; JSTOR Archive Collection A-Z Listing; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Animals Biological and medical sciences Cell Culture Techniques Cell Line, Tumor Cell lines Cryptosporidiosis Cryptosporidiosis - parasitology Cryptosporidiosis - pathology Cryptosporidium parvum Cryptosporidium parvum - physiology Epithelial cells Epithelial Cells - parasitology Epithelial Cells - pathology Fundamental and applied biological sciences. Psychology Human protozoal diseases Humans Infections Infectious diseases Medical sciences Microbiology Microgravity Microvilli Oocysts Organoids Organoids - cytology Organoids - parasitology Parasites Parasitic diseases Polymerase chain reaction Protozoal diseases Rotation Vicia villosa |
| title | Detection of Epithelial-Cell Injury, and Quantification of Infection, in the HCT-8 Organoid Model of Cryptosporidiosis |
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