Establishment and Characterization of Canine Mammary Gland Carcinoma Cell Lines with Vasculogenic Mimicry Ability <em>in vitro</em> and <em>in vivo</em>

Abstract

Mammary neoplasms affect a population of uncastrated and elderly female dogs and most of these neoplasms are malignant. In order to study this disease cell culture presents itself as a promising preclinical model, creating the opportunity to deposit cell lines at a cell bank, allowing a great repetition of the assays and making the validation of the results more reliable. Including, in vitro experiments for vasculogenic mimicry (VM) evaluation. VM is related to cancer cells capable of generate vascular-like structures without endothelial cells, mimicking the vasculogenic process. The aim of this study was to establish and characterize ten cell lines from canine mammary gland tumour according to immunophenotype and tumorigenicity, and with its ability to form vasculogenic mimicry-like structures in vitro. Fifteen samples from canine mammary gland carcinoma were collected and cultured in vitro and ten cell lines were established and characterized. Cells were evaluated for morphology, phenotype, vascular mimicry and tumorigenicity. All cell lines presented spindle shape morphology and expressed concomitant pan-cytokeratin and CK8/18. Four cell lines had vasculogenic mimicry ability and two cell showed in vivo tumorigenic potential. Cell characterization of those lines will help to create a database for more knowledge of mammary gland carcinomas in dogs, including studies of tumor behavior and new therapeutic targets.

Effects of Lapatinib on HER2-Positive and HER2-Negative Canine Mammary Carcinoma Cells Cultured In Vitro

“…Samples that met all of the following criteria were included in this research: cell cultures of primary tumors that had paired formalin-fixed paraffin-embedded (FFPE) material and were classified according to Goldschmidt et al [ 29 ], cell cultures characterized by cell phenotype and tumorigenicity in vitro, cell cultures with sufficient aliquots for triplicate analysis in all experiments and free of bacterial, mycoplasma and/or fungal contamination. Cell lines were previously established and characterized [ 30 ]. A total of six cell lines met the criteria.…”

“…The cell culture UNESP-CM1 was used on the 15th passage, UNESP-CM5 on the 16th, UNESP-CM9 on the 17th, UNESP-CM60 on the 35th, UNESP-MM1 on the 19th, and UNESP-MM4 on the 22nd. The passages used were chosen according to cell stability and aliquot availability [ 30 ].…”

“…The drug was diluted in DMSO to a concentration of 2 mg/mL, and the highest dilution was used as the parameter for the control containing DMSO (at the same concentration as the highest concentration of lapatinib). Cells were cultured according to Lainetti et al [ 30 ].…”

Investigation of the Prognostic Significance of Vasculogenic Mimicry and Its Inhibition by Sorafenib in Canine Mammary Gland Tumors

“…The establishment of canine mammary cell cultures followed the previous description published by our research group (21), and all procedures for the establishment, characterization and culture of CM9 and CM60 mammary primary cells were described previously (22). The anti-tumoural effect of sorafenib was determined by an assay based on the cleavage of an MTT salt into purple crystals by metabolically active cells.…”