Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property

Abstract

Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine. Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model. Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR+, CD5.1+, CD14+, CD45+, CD83+, CD163+, and Ly-6G-Ly-6C+. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×108 did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells. Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.

Diagnose Enforcement of Case Canine Transmissible Venereal Tumor (CTVT) on Domestic Dog

“…Results of CTVT histopathological examination in Domestic Dogs from vulvavagina biopsy showed the presence of infiltration of perivascular inflammatory cells, neovascularization, and proliferation of fibroblast cells accompanied by necrosis and the formation of cytoplasmic vacuoles. The results of this examination are in accordance with the results of research by Zayas et al [22], which suggests that the proliferation of fibroblast cells indicates the expansion of tumor cells in tissues or organs. Fibroblasts or connective tissues have a function as a supporting framework and binders of parenchymal neoplasmic cells forming tissue masses.…”

In vitro chemosensitivity of a canine tumor venereal transmissible cancer cell line

“…For maintenance, CTVT cells stablished by our laboratory ( 2 ) were cultured in a 75 cm 2 culture flask (Corning, USA) containing Dulbecco's modified Eagle's medium (DMEM/F-12) and 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES) (Gibco; Thermo Fisher Scientific, USA) with 10% fetal bovine serum (FBS) (Gibco), as well as antibiotics and antimycotics (Antibiotic-Antimycotic 100X; Gibco), at 37 °C with 5% CO 2 . StemPro Accutase (Gibco) was used to detach the cells from the flask ( 2 ).…”

“…The canine transmissible venereal tumor (CTVT) is a neoplasia transmitted sexually by the transplantation of viable tumor cells; although it mostly occurs in the genital region, implantation can occur in any part of the body, especially if there are surface abrasions or loss of integrity of the skin ( 1 , 2 ). Vincristine sulfate is a commonly used chemotherapeutic to treat this neoplasm, reporting a high degree of efficacious and complete remission in more than 90% of the cases.…”

“…However, there are clinical reports that mention vincristine sulfate resistance in CTVT ( 3 ). The chemotherapeutic options to be used in the case of CTVT treatment or resistance are limited due to the scarce clinical and in vitro chemo sensibility studies that suggest new options; one of the reasons that for lack of studies is that no CTVT cell line exists except for the one used in this study ( 2 ). Today, studies in patients and in vitro experiments using cell cultures are used to analyze the response to drug treatments.…”

Between the Devil and the Deep Blue Sea: Non-Coding RNAs Associated with Transmissible Cancers in Tasmanian Devil, Domestic Dog and Bivalves

“…CRISPR/Cas screens have been performed to identify the functional roles of lncRNAs in human-derived cancer cell lines [ 120 , 121 ], but have not been carried out for transmissible cancers. Although no stable cell lines currently exist for BTN, there are cell lines readily available for DFTD and CTVT [ 122 ]. These provide the possibility for CRISPR/Cas screens to start disentangling the lncRNA biology that contributes to the proliferation and transmissibility of tumours.…”