Ultrasound-guided intramural inoculation of orthotopic bladder cancer xenografts: a novel high-precision approach
Orthotopic bladder cancer xenografts are essential for testing novel therapies and molecular manipulations of cell lines in vivo. Current xenografts rely on tumor cell inoculation by intravesical instillation or direct injection into the bladder wall. Instillation is limited by the lack of cell line...
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| creator | Jäger, Wolfgang Moskalev, Igor Janssen, Claudia Hayashi, Tetsutaro Awrey, Shannon Gust, Kilian M So, Alan I Zhang, Kaixin Fazli, Ladan Li, Estelle Thüroff, Joachim W Lange, Dirk Black, Peter C |
| description | Orthotopic bladder cancer xenografts are essential for testing novel therapies and molecular manipulations of cell lines in vivo. Current xenografts rely on tumor cell inoculation by intravesical instillation or direct injection into the bladder wall. Instillation is limited by the lack of cell lines that are tumorigenic when delivered in this manner. The invasive model inflicts morbidity on the mice by the need for laparotomy and mobilization of the bladder. Furthermore this procedure is complex and time-consuming. Three bladder cancer cell lines (UM-UC1, UM-UC3, UM-UC13) were inoculated into 50 athymic nude mice by percutaneous injection under ultrasound guidance. PBS was first injected between the muscle wall and the mucosa to separate these layers, and tumor cells were subsequently injected into this space. Bioluminescence and ultrasound were used to monitor tumor growth. Contrast-enhanced ultrasound was used to study changes in tumor perfusion after systemic gemcitabine/cisplatin treatment. To demonstrate proof of principle that therapeutic agents can be injected into established xenografts under ultrasound guidance, oncolytic virus (VSV) was injected into UM-UC3 tumors. Xenograft tissue was harvested for immunohistochemistry after 23-37 days. Percutaneous injection of tumor cells into the bladder wall was performed efficiently (mean time: 5.7 min) and without complications in all 50 animals. Ultrasound and bioluminescence confirmed presence of tumor in the anterior bladder wall in all animals 3 days later. The average tumor volumes increased steadily over the study period. UM-UC13 tumors showed a marked decrease in volume and perfusion after chemotherapy. Immunohistochemical staining for VSV-G demonstrated virus uptake in all UM-UC3 tumors after intratumoral injection. We have developed a novel method for creating orthotopic bladder cancer xenograft in a minimally invasive fashion. In our hands this has replaced the traditional model requiring laparotomy, because this model is more time efficient, more precise and associated with less morbidity for the mice. |
| doi_str_mv | 10.1371/journal.pone.0059536 |
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Current xenografts rely on tumor cell inoculation by intravesical instillation or direct injection into the bladder wall. Instillation is limited by the lack of cell lines that are tumorigenic when delivered in this manner. The invasive model inflicts morbidity on the mice by the need for laparotomy and mobilization of the bladder. Furthermore this procedure is complex and time-consuming. Three bladder cancer cell lines (UM-UC1, UM-UC3, UM-UC13) were inoculated into 50 athymic nude mice by percutaneous injection under ultrasound guidance. PBS was first injected between the muscle wall and the mucosa to separate these layers, and tumor cells were subsequently injected into this space. Bioluminescence and ultrasound were used to monitor tumor growth. Contrast-enhanced ultrasound was used to study changes in tumor perfusion after systemic gemcitabine/cisplatin treatment. To demonstrate proof of principle that therapeutic agents can be injected into established xenografts under ultrasound guidance, oncolytic virus (VSV) was injected into UM-UC3 tumors. Xenograft tissue was harvested for immunohistochemistry after 23-37 days. Percutaneous injection of tumor cells into the bladder wall was performed efficiently (mean time: 5.7 min) and without complications in all 50 animals. Ultrasound and bioluminescence confirmed presence of tumor in the anterior bladder wall in all animals 3 days later. The average tumor volumes increased steadily over the study period. UM-UC13 tumors showed a marked decrease in volume and perfusion after chemotherapy. Immunohistochemical staining for VSV-G demonstrated virus uptake in all UM-UC3 tumors after intratumoral injection. We have developed a novel method for creating orthotopic bladder cancer xenograft in a minimally invasive fashion. In our hands this has replaced the traditional model requiring laparotomy, because this model is more time efficient, more precise and associated with less morbidity for the mice.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0059536</identifier><identifier>PMID: 23555699</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adenoviruses ; Angiogenesis ; Animals ; Biology ; Bioluminescence ; Biotechnology ; Bladder ; Bladder cancer ; Cancer ; Cell Line, Tumor ; Cell Proliferation ; Chemical compounds ; Chemotherapy ; Cisplatin ; Complications ; Feasibility Studies ; Female ; Gemcitabine ; Gene therapy ; Humans ; Immunohistochemistry ; In vivo methods and tests ; Injection ; Inoculation ; Medicine ; Mice ; Morbidity ; Mucosa ; Muscles ; NMR ; Nuclear magnetic resonance ; Oncolysis ; Oncolytic Viruses - physiology ; Perfusion ; Pharmacology ; Prostate ; Surgery, Computer-Assisted - methods ; Transplantation, Heterologous - methods ; Tumor cell lines ; Tumor cells ; Tumors ; Ultrasonic imaging ; Ultrasonics ; Ultrasound ; Urinary bladder ; Urinary Bladder Neoplasms - drug therapy ; Urinary Bladder Neoplasms - pathology ; Urinary Bladder Neoplasms - virology ; Urology ; Vascular endothelial growth factor ; Viruses ; Xenografts ; Xenotransplantation</subject><ispartof>PloS one, 2013-03, Vol.8 (3), p.e59536-e59536</ispartof><rights>2013 Jäger et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Jäger et al 2013 Jäger et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-835a024c5c825bf3b079d89b6225d89eb30b49970bcb12877d9b1426f17b8c03</citedby><cites>FETCH-LOGICAL-c526t-835a024c5c825bf3b079d89b6225d89eb30b49970bcb12877d9b1426f17b8c03</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/PMC3608695/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3608695/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23555699$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Zi, Xiaolin</contributor><creatorcontrib>Jäger, Wolfgang</creatorcontrib><creatorcontrib>Moskalev, Igor</creatorcontrib><creatorcontrib>Janssen, Claudia</creatorcontrib><creatorcontrib>Hayashi, Tetsutaro</creatorcontrib><creatorcontrib>Awrey, Shannon</creatorcontrib><creatorcontrib>Gust, Kilian M</creatorcontrib><creatorcontrib>So, Alan I</creatorcontrib><creatorcontrib>Zhang, Kaixin</creatorcontrib><creatorcontrib>Fazli, Ladan</creatorcontrib><creatorcontrib>Li, Estelle</creatorcontrib><creatorcontrib>Thüroff, Joachim W</creatorcontrib><creatorcontrib>Lange, Dirk</creatorcontrib><creatorcontrib>Black, Peter C</creatorcontrib><title>Ultrasound-guided intramural inoculation of orthotopic bladder cancer xenografts: a novel high-precision approach</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Orthotopic bladder cancer xenografts are essential for testing novel therapies and molecular manipulations of cell lines in vivo. Current xenografts rely on tumor cell inoculation by intravesical instillation or direct injection into the bladder wall. Instillation is limited by the lack of cell lines that are tumorigenic when delivered in this manner. The invasive model inflicts morbidity on the mice by the need for laparotomy and mobilization of the bladder. Furthermore this procedure is complex and time-consuming. Three bladder cancer cell lines (UM-UC1, UM-UC3, UM-UC13) were inoculated into 50 athymic nude mice by percutaneous injection under ultrasound guidance. PBS was first injected between the muscle wall and the mucosa to separate these layers, and tumor cells were subsequently injected into this space. Bioluminescence and ultrasound were used to monitor tumor growth. Contrast-enhanced ultrasound was used to study changes in tumor perfusion after systemic gemcitabine/cisplatin treatment. To demonstrate proof of principle that therapeutic agents can be injected into established xenografts under ultrasound guidance, oncolytic virus (VSV) was injected into UM-UC3 tumors. Xenograft tissue was harvested for immunohistochemistry after 23-37 days. Percutaneous injection of tumor cells into the bladder wall was performed efficiently (mean time: 5.7 min) and without complications in all 50 animals. Ultrasound and bioluminescence confirmed presence of tumor in the anterior bladder wall in all animals 3 days later. The average tumor volumes increased steadily over the study period. UM-UC13 tumors showed a marked decrease in volume and perfusion after chemotherapy. Immunohistochemical staining for VSV-G demonstrated virus uptake in all UM-UC3 tumors after intratumoral injection. We have developed a novel method for creating orthotopic bladder cancer xenograft in a minimally invasive fashion. In our hands this has replaced the traditional model requiring laparotomy, because this model is more time efficient, more precise and associated with less morbidity for the mice.</description><subject>Adenoviruses</subject><subject>Angiogenesis</subject><subject>Animals</subject><subject>Biology</subject><subject>Bioluminescence</subject><subject>Biotechnology</subject><subject>Bladder</subject><subject>Bladder cancer</subject><subject>Cancer</subject><subject>Cell Line, Tumor</subject><subject>Cell Proliferation</subject><subject>Chemical compounds</subject><subject>Chemotherapy</subject><subject>Cisplatin</subject><subject>Complications</subject><subject>Feasibility Studies</subject><subject>Female</subject><subject>Gemcitabine</subject><subject>Gene therapy</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>In vivo methods and tests</subject><subject>Injection</subject><subject>Inoculation</subject><subject>Medicine</subject><subject>Mice</subject><subject>Morbidity</subject><subject>Mucosa</subject><subject>Muscles</subject><subject>NMR</subject><subject>Nuclear magnetic resonance</subject><subject>Oncolysis</subject><subject>Oncolytic Viruses - physiology</subject><subject>Perfusion</subject><subject>Pharmacology</subject><subject>Prostate</subject><subject>Surgery, Computer-Assisted - methods</subject><subject>Transplantation, Heterologous - methods</subject><subject>Tumor cell lines</subject><subject>Tumor cells</subject><subject>Tumors</subject><subject>Ultrasonic imaging</subject><subject>Ultrasonics</subject><subject>Ultrasound</subject><subject>Urinary bladder</subject><subject>Urinary Bladder Neoplasms - drug therapy</subject><subject>Urinary Bladder Neoplasms - pathology</subject><subject>Urinary Bladder Neoplasms - virology</subject><subject>Urology</subject><subject>Vascular endothelial growth 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C</au><au>Zi, Xiaolin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ultrasound-guided intramural inoculation of orthotopic bladder cancer xenografts: a novel high-precision approach</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-03-26</date><risdate>2013</risdate><volume>8</volume><issue>3</issue><spage>e59536</spage><epage>e59536</epage><pages>e59536-e59536</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Orthotopic bladder cancer xenografts are essential for testing novel therapies and molecular manipulations of cell lines in vivo. Current xenografts rely on tumor cell inoculation by intravesical instillation or direct injection into the bladder wall. Instillation is limited by the lack of cell lines that are tumorigenic when delivered in this manner. The invasive model inflicts morbidity on the mice by the need for laparotomy and mobilization of the bladder. Furthermore this procedure is complex and time-consuming. Three bladder cancer cell lines (UM-UC1, UM-UC3, UM-UC13) were inoculated into 50 athymic nude mice by percutaneous injection under ultrasound guidance. PBS was first injected between the muscle wall and the mucosa to separate these layers, and tumor cells were subsequently injected into this space. Bioluminescence and ultrasound were used to monitor tumor growth. Contrast-enhanced ultrasound was used to study changes in tumor perfusion after systemic gemcitabine/cisplatin treatment. To demonstrate proof of principle that therapeutic agents can be injected into established xenografts under ultrasound guidance, oncolytic virus (VSV) was injected into UM-UC3 tumors. Xenograft tissue was harvested for immunohistochemistry after 23-37 days. Percutaneous injection of tumor cells into the bladder wall was performed efficiently (mean time: 5.7 min) and without complications in all 50 animals. Ultrasound and bioluminescence confirmed presence of tumor in the anterior bladder wall in all animals 3 days later. The average tumor volumes increased steadily over the study period. UM-UC13 tumors showed a marked decrease in volume and perfusion after chemotherapy. Immunohistochemical staining for VSV-G demonstrated virus uptake in all UM-UC3 tumors after intratumoral injection. We have developed a novel method for creating orthotopic bladder cancer xenograft in a minimally invasive fashion. In our hands this has replaced the traditional model requiring laparotomy, because this model is more time efficient, more precise and associated with less morbidity for the mice.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23555699</pmid><doi>10.1371/journal.pone.0059536</doi><oa>free_for_read</oa></addata></record> |
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| ispartof | PloS one, 2013-03, Vol.8 (3), p.e59536-e59536 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1330899156 |
| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adenoviruses Angiogenesis Animals Biology Bioluminescence Biotechnology Bladder Bladder cancer Cancer Cell Line, Tumor Cell Proliferation Chemical compounds Chemotherapy Cisplatin Complications Feasibility Studies Female Gemcitabine Gene therapy Humans Immunohistochemistry In vivo methods and tests Injection Inoculation Medicine Mice Morbidity Mucosa Muscles NMR Nuclear magnetic resonance Oncolysis Oncolytic Viruses - physiology Perfusion Pharmacology Prostate Surgery, Computer-Assisted - methods Transplantation, Heterologous - methods Tumor cell lines Tumor cells Tumors Ultrasonic imaging Ultrasonics Ultrasound Urinary bladder Urinary Bladder Neoplasms - drug therapy Urinary Bladder Neoplasms - pathology Urinary Bladder Neoplasms - virology Urology Vascular endothelial growth factor Viruses Xenografts Xenotransplantation |
| title | Ultrasound-guided intramural inoculation of orthotopic bladder cancer xenografts: a novel high-precision approach |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-30T07%3A08%3A53IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Ultrasound-guided%20intramural%20inoculation%20of%20orthotopic%20bladder%20cancer%20xenografts:%20a%20novel%20high-precision%20approach&rft.jtitle=PloS%20one&rft.au=J%C3%A4ger,%20Wolfgang&rft.date=2013-03-26&rft.volume=8&rft.issue=3&rft.spage=e59536&rft.epage=e59536&rft.pages=e59536-e59536&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0059536&rft_dat=%3Cproquest_plos_%3E2949868801%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1330899156&rft_id=info:pmid/23555699&rft_doaj_id=oai_doaj_org_article_ebcb698bc91941febf61917be534a7b3&rfr_iscdi=true |