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Establishment and Partial Characterization of Canine Mammary Tumor Cell Lines. Animals 2025, 15, 1991. [https://doi.org/](https://doi.org/)[ ](https://doi.org/)[10.3390/ani15131991](10.3390/ani15131991)  \nCopyright: © 2025 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://creativecommons.org/](https://creativecommons.org/)[ ](https://creativecommons.org/)[licenses/by/4.0/](licenses/by/4.0/)) .  \n1 Department of Chemistry-Biology, University of Sonora, Blvd. Luis Encinas y Rosales s/n, Hermosillo 83000, Mexico; [eliza.vazquez@unison.mx](eliza.vazquez@unison.mx) (E.V.); [a214206530@unison.mx](a214206530@unison.mx) (L.D.); [a217217239@unison.mx](a217217239@unison.mx) (B.S.); [a210206416@unison.mx](a210206416@unison.mx) (G.T.); [adriana.garibay@unison.mx](adriana.garibay@unison.mx) (A.G.-E.)  \n2 Veterinary and Animal Research Centre (CECAV), University of Trás-os-Montes and Alto Douro, 5001-801 Vila Real, Portugal  \n* Correspondence: [fqueirog@utad.pt](fqueirog@utad.pt) (F.L.Q.); [velaz2@unison.mx](velaz2@unison.mx) (C.V.)  \nSimple Summary  \nMammary tumors are the most frequent neoplasia in female dogs. Canine mammary cell lines represent a model of study for women breast cancer research that could benefit both species. The aim of our study was to establish and partially characterize canine mammary tumor cell lines. Ten cell cultures were generated from tumor tissue obtained from canines with mammary tumors. Seven cell lines were generated from primary mammary tumorsand three cell lines from metastatic sites. Immunocytochemistry (ICC) analysis revealed a negative expression of hormonal receptors (estrogen receptor [ER] and progesterone receptor [PR]) in five cell lines and positive in one cell line, whereas six cell lines were human epidermal growth factor receptor 2 (HER2)-negative and positive for vimentin. Several cell lines showed tumorigenicity capacity in vitro. The susceptibility of five cell lines to different drugs was evaluated with the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, where doxorubicin (DOX) showed the highest growth-inhibitory effect (DOX > Paclitaxel > Colchicine > 5-Fluorouracil [5-FU] > Carboplatin) . The cell lines generated represent a model of study for breast cancer that can be used for developing and testing new treatments to improve the survival of canine and human patients.  \nAbstract  \nMammary tumors are the most common neoplasms diagnosed in female dogs and have been considered excellent models for studying human breast cancer. Establishing cell lines from primary cultures of canine mammary tumors provides an in vitro model to better understand the disease and develop new treatments. This study aimed to establish and characterize canine mammary tumor cell lines. Ten cell cultures were generated from tumor tissue obtained from affected dogs, including seven from primary mammary tumorsand three from metastatic sites. Characterization included molecular marker expression (ER, PR, HER2, cytokeratin 5/6 (CK5/6), vimentin, and the marker of cell proliferation Ki67) and in vitro tumorigenic capacity assessment. Additionally, the susceptibility of five cell lines to DOX, 5-FU, paclitaxel, colchicine, and carboplatin was evaluated using the MTT assay. ICC analysis revealed negative expression of","cbCailYsxR0df6xI","https://ap.wps.com/l/cbCailYsxR0df6xI","pdf",25775412,"English","# Simple Summary\n# Abstract\n# Keywords\n# Introduction","[{\"question\":\"How were the canine mammary tumor cell lines established in the study?\",\"answer\":\"Ten cell cultures were generated from tumor tissue collected from dogs with mammary tumors, including seven derived from primary tumors and three from metastatic sites.\"},{\"question\":\"Which molecular markers were evaluated during characterization?\",\"answer\":\"The study examined ER, PR, HER2, CK5/6, vimentin, and the proliferation marker Ki67 using immunocytochemistry and related analyses.\"},{\"question\":\"What did the MTT assay show about drug sensitivity of the cell lines?\",\"answer\":\"Using the MTT assay, doxorubicin produced the highest growth-inhibitory effect among tested drugs, following an order of effectiveness described in the results. Some lines were resistant to carboplatin and 5-FU.\"}]","Establishment and Partial Characterization of Canine Mammary Tumor Cell Lines | PDF",1790202016,48]