{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:24:20Z","timestamp":1760149460480,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2023,8,16]],"date-time":"2023-08-16T00:00:00Z","timestamp":1692144000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Portuguese Foundation for Science and Technology (FCT), Portugal","award":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"],"award-info":[{"award-number":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"]}]},{"name":"Pfizer","award":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"],"award-info":[{"award-number":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"]}]},{"name":"Magyar Tudom\u00e1nyos Akad\u00e9mia","award":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"],"award-info":[{"award-number":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"]}]},{"name":"National Research Development and Innovation Office (NKFIH), Hungary","award":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"],"award-info":[{"award-number":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"]}]},{"name":"Executive Agency for Higher Education, Research, Development and Innovation Funding (UEFISCDI), Romania","award":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"],"award-info":[{"award-number":["FCT-PTD\/MED-ONC\/29402\/2017","UIDB\/04138\/2020","UIDP\/04138\/2020","SFRH\/BD\/145522\/2019","68977633","NAP2022-I-7\/2022","K135425","TKP2021-EGA-09","PN-III-P4-ID-PCE-2020-1529"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Breast cancer (BC) brain metastases (BCBM) is a severe condition frequently occurring in the triple-negative subtype. The study of BCBM pathogenesis and treatment has been hampered by the difficulty in establishing a reliable animal model that faithfully recapitulates the preferential formation of brain metastases. The injection of BC cells in the carotid artery of mice has been proposed but the procedure is challenging, with the metastatic pattern being scarcely characterized. In this work, we thoroughly describe an improved procedure, highlighting the tricks and challenges of the process, and providing a characterization of the brain and peripheral metastatic pattern at the cellular and molecular level. Triple-negative BC (4T1) cells were inoculated in the common carotid artery of BALB\/c mice. Brains and peripheral organs were harvested at 7\u201314 days for the histological characterization of the metastases\u2019 pattern and the immunofluorescence analysis of specific markers. With our surgical procedure, both mouse death and procedure-associated weight loss were negligible. Brain metastases mostly occurred in the hippocampus, while sparse peripheral lesions were only detected in the lungs. Brain-colonizing BC cells presented proliferative (Ki-67) and epithelial (pan-cytokeratin and tomato lectin) features, which account for metastases\u2019 establishment. The presented surgical approach constitutes an important and reliable tool for BCBM studies.<\/jats:p>","DOI":"10.3390\/cells12162076","type":"journal-article","created":{"date-parts":[[2023,8,16]],"date-time":"2023-08-16T09:57:02Z","timestamp":1692179822000},"page":"2076","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Breast Cancer Brain Metastases: Implementation and Characterization of a Mouse Model Relying on Malignant Cells Inoculation in the Carotid Artery"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8219-6240","authenticated-orcid":false,"given":"Joana","family":"Godinho-Pereira","sequence":"first","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmaceutical Sciences and Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"given":"Daniela","family":"Vaz","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmaceutical Sciences and Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4639-6883","authenticated-orcid":false,"given":"In\u00eas","family":"Figueira","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Farm-ID\u2014Faculty of Pharmacy Research and Development Association, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"given":"Joana","family":"Aniceto-Rom\u00e3o","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmaceutical Sciences and Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"given":"Istvan","family":"Krizbai","sequence":"additional","affiliation":[{"name":"Institute of Biophysics, Biological Research Centre, E\u00f6tv\u00f6s Lor\u00e1nd Research Network (ELKH), 6726 Szeged, Hungary"},{"name":"Institute of Life Sciences, Vasile Goldis Western University of Arad, Str. Liviu Rebreanu 86, 310414 Arad, Romania"}]},{"given":"Rui","family":"Malh\u00f3","sequence":"additional","affiliation":[{"name":"BioISI\u2014Biosystems and Integrative Sciences Institute, Faculty of Sciences, Universidade de Lisboa, Campo Grande, 1746-016 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0303-8085","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Rocha","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmacy, Pharmacology and Health Technologies, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6028-4717","authenticated-orcid":false,"given":"Manuela Colla","family":"Carvalheiro","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmacy, Pharmacology and Health Technologies, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"given":"Sandra","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmacy, Pharmacology and Health Technologies, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6814-7226","authenticated-orcid":false,"given":"Maria Manuela","family":"Gaspar","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmacy, Pharmacology and Health Technologies, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8493-4649","authenticated-orcid":false,"given":"Maria Alexandra","family":"Brito","sequence":"additional","affiliation":[{"name":"iMed.ULisboa\u2014Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"},{"name":"Department of Pharmaceutical Sciences and Medicines, Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,8,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"363","DOI":"10.3322\/caac.21565","article-title":"Cancer treatment and survivorship statistics, 2019","volume":"69","author":"Miller","year":"2019","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41416-020-01161-4","article-title":"The lingering mysteries of metastatic recurrence in breast cancer","volume":"124","author":"Riggio","year":"2021","journal-title":"Br. J. 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