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A common in vivo approach injects each cell type into animals, but it demands many subjects and months to yield meaningful results. This work introduces a method using cancer spheroids embedded in tissue-engineered skin substitutes (cTES), grafted onto athymic mice, and compares it with a cell injection assay. Tumors formed in both models, while metastases were not observed at sacrifice.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/cancer-spheroids-embedded-in-tissue-engineered-skin-substitutes-a-new-method-to-study-tumorigenicity-in-vivo/330084/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/cancer-spheroids-embedded-in-tissue-engineered-skin-substitutes-a-new-method-to-study-tumorigenicity-in-vivo/330084.png","ImageObject",300,407,{"name":92,"@type":93},"Himbo","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-11-04","2026-09-21",true,{"@type":102,"interactionType":103,"userInteractionCount":39},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What challenge do traditional in vivo tumorigenic assays present for clinical translation?","Question",{"text":112,"@type":113},"They typically require many animals and several months to obtain useful results, making them time-consuming, costly, and ethically concerning.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How does the proposed method study tumorigenicity?",{"text":117,"@type":113},"It embeds cancer spheroids into tissue-engineered skin substitutes (cTES), grafts them onto athymic mice, and compares outcomes with a commonly used cell injection assay.",{"name":119,"@type":110,"acceptedAnswer":120},"What were the observed differences between the two tumorigenicity models?",{"text":121,"@type":113},"Both models developed tumors, no metastases were detected at sacrifice, and the cTES approach showed a faster tumor development rate than the injection method.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},330084,1790575436,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":39,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145,"preview_page_count":4},687197100911,"https://ap-avatar.wpscdn.com/avatar/a000239b6f1da00475?x-image-process=image/resize,m_fixed,w_180,h_180&k=1785132997149421697","Article  \nCancer Spheroids Embedded in Tissue-Engineered Skin Substitutes: A New Method to Study Tumorigenicity In Vivo  \nMartin A. Barbier 1,2,3, Karel Ferland 1,2,3, Henri De Koninck 1,2,3, Emilie J. Doucet 1,2,3,  \nLudivine Dubourget 1,2,3, MinJoon Kim 1,2,3, Bettina Cattier 1,2,3, Amélie Morissette 1,3, Mbarka Bchetnia 1,3, Danielle Larouche 1,3, Dong Hyun Kim 1,3,4, Guillaume St-Jean 5 and Lucie Germain 1,2,3, *  \nCitation: Barbier, M.A.; Ferland, K.; De Koninck, H.; Doucet, E.J.;  \nDubourget, L.; Kim, M.; Cattier, B.; Morissette, A.; Bchetnia, M.; Larouche, D.; et al. Cancer Spheroids Embedded in Tissue-Engineered Skin Substitutes: ANew Method to Study Tumorigenicity In Vivo. Int. J. Mol. Sci. 2024, 25, 1513 . [https://doi.org/10.3390/](https://doi.org/10.3390/)[ ](https://doi.org/10.3390/)ijms25031513  \nAcademic Editor: Agnes Klar  \nReceived: 28 December 2023  \nRevised: 20 January 2024  \nAccepted: 23 January 2024  \nPublished: 26 January 2024  \nCopyright: © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) .  \n1 The Tissue Engineering Laboratory (LOEX), Université Laval’s Research Center, Quebec, QC G1V 0A6, Canada; [martin-alexandre.barbier.1@ulaval.ca](martin-alexandre.barbier.1@ulaval.ca) (M.A.B.);  \n[karel.ferland.1@ulaval.ca](karel.ferland.1@ulaval.ca) (K.F.); [henri.de-koninck.1@ulaval.ca](henri.de-koninck.1@ulaval.ca) (H.D.K.); [emilie.doucet.4@ulaval.ca](emilie.doucet.4@ulaval.ca) (E.J.D.);  \n[ludivine.dubourget.1@ulaval.ca](ludivine.dubourget.1@ulaval.ca) (L.D.); [mjk042821@gmail.com](mjk042821@gmail.com) (M.K.); bettina.cattier.1@ulaval.ca (B.C.);  \n[amelie.morissette@crchudequebec.ulaval.ca](amelie.morissette@crchudequebec.ulaval.ca) (A.M.); [mbarka.bchetnia@crchudequebec.ulaval.ca](mbarka.bchetnia@crchudequebec.ulaval.ca) (M.B.);  \n[danielle.larouche@crchudequebec.ulaval.ca](danielle.larouche@crchudequebec.ulaval.ca) (D.L.); [terios92@hanmail.net](terios92@hanmail.net) (D.H.K.)  \n2 Department of Surgery, Faculty of Medicine, Université Laval, Quebec, QC G1V 0A6, Canada  \n3 Regenerative Medicine Division, CHU de Québec-Université Laval Research Centre, Quebec, QC G1J 1Z4, Canada  \n4 Department of Dermatology, CHA Bundang Medical Center, CHA University, 59 Yatap-ro, Bundang-gu, Seongnam-si 463-712, Gyeonggi-do, Republic of Korea  \n5 Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, QC J2S 2M2, Canada; [guillaume.st-jean.2@umontreal.ca](guillaume.st-jean.2@umontreal.ca)  \n* Correspondence: [lucie.germain@fmed.ulaval.ca](lucie.germain@fmed.ulaval.ca); Tel.: +1-418-525-4444 (ext. 61696)  \nAbstract: Tumorigenic assays are used during a clinical translation to detect the transformation potential of cell-based therapies. One of these in vivo assays is based on the separate injection of each cell type to be used in the clinical trial. However, the injection method requires many animals and several months to obtain useful results. In previous studies, we showed the potential of tissueengineered skin substitutes (TESs) as a model for normal skin in which cancer cells can be included in vitro. Herein, we showed a new method to study tumorigenicity, using cancer spheroids that were embedded in TESs (cTES) and grafted onto athymic mice, and compared it with the commonly used cell injection assay. Tumors developed in both models, cancer cell injection and cTES grafting, but metastases were not detected at the time of sacrifice. Interestingly, the rate of tumor development was faster in cTESs than with the injection method. In conclusion, grafting TESs is a sensitive method to detect tumor cell growth with and could be developed as an alternative test for tumorigenicity.  \n","cbCaicRagyFU3Cn5","https://ap.wps.com/l/cbCaicRagyFU3Cn5","pdf",5581627,14,"English","# Introduction\n## Rationale and limitations of in vivo tumorigenic assays\n## Common subcutaneous injection approach and validation criteria","[{\"question\":\"What challenge do traditional in vivo tumorigenic assays present for clinical translation?\",\"answer\":\"They typically require many animals and several months to obtain useful results, making them time-consuming, costly, and ethically concerning.\"},{\"question\":\"How does the proposed method study tumorigenicity?\",\"answer\":\"It embeds cancer spheroids into tissue-engineered skin substitutes (cTES), grafts them onto athymic mice, and compares outcomes with a commonly used cell injection assay.\"},{\"question\":\"What were the observed differences between the two tumorigenicity models?\",\"answer\":\"Both models developed tumors, no metastases were detected at sacrifice, and the cTES approach showed a faster tumor development rate than the injection method.\"}]","Cancer Spheroids Embedded in Tissue-Engineered Skin Substitutes - A New Method to Study Tumorigenicity In Vivo | PDF",1789987045,35]