Cellular and Phenotypic Characterization of Canine Osteosarcoma Cell Lines

Abstract

Canine and human osteosarcoma (OSA) have many similarities, with the majority of reported cases occurring in the appendicular skeleton, gender predominance noted, high rate of metastasis at the time of presentation, and a lack of known etiology for this devastating disease. Due to poor understanding of the molecular mechanisms underlying OSA, we have characterized seven different OSA canine cell lines: Abrams, D17, Grey, Hughes, Ingles, Jarques, and Marisco and compared them to U2, a human OSA cell line, for the following parameters: morphology, growth, contact inhibition, migrational tendencies, alkaline phosphatase staining, heterologous tumor growth, double-strand DNA breaks, and oxidative damage. All results demonstrated the positive characteristics of the Abrams cell line for use in future studies of OSA. Of particular interest, the robust growth of a subcutaneous tumor and rapid pulmonary metastasis of the Abrams cell line in an immunocompromised mouse shows incredible potential for the future use of Abrams as a canine OSA model. Further investigations utilizing a canine cell model of OSA, such as Abrams, will be invaluable to understanding the molecular events underlying OSA, pharmaceutical inhibition of metastasis, and eventual prevention of this devastating disease.

Establishment of three‐dimensional canine osteosarcoma cell lines showing vasculogenic mimicry and evaluation of biological properties after treatment with 17‐AAG

“…These different cellular properties could be explained by the invasiveness and aggressive behaviour of D17 cell lines, derived from a metastatic lesion, as well as by the cellular phenotype acquired during the growth passages. 30 Histological investigation of D17 3D long-term cultures further showed the presence of vascular-like channels showing cavities in serial transversal sections surrounded by tumour endothelial-like cells identifiable using immunohistochemistry. As shown in results, the lack of CD31 expression excluded the ability of D17 cells to completely differentiate into endothelium in 3D cultures.…”

“…Our study was firstly focused on two canine OSA cell lines (D17 and D22), revealing that D17 cell line showed a greater ability to grow in different conditions in comparison to D22 cell line or fibroblasts. These different cellular properties could be explained by the invasiveness and aggressive behaviour of D17 cell lines, derived from a metastatic lesion, as well as by the cellular phenotype acquired during the growth passages …”

Relative biological effectiveness in canine osteosarcoma cells irradiated with accelerated charged particles

“…The canine OSA cell lines Abrams, D17, Grey and Moresco were a gift from the Animal Cancer Center of Colorado State University (Fort Collins, CO, USA) (22,24). Cells were grown in minimal essential media (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% (v/v) Pen/Strep and fungizone solution (Invitrogen; Thermo Fisher Scientific, Inc.), and they were maintained in a tissue culture incubator at 37˚C in a 100% humidified atmosphere of 5% CO 2 .…”

DNA damage response and DNA repair – dog as a model?

“…Thus suggesting high homology between the proteins of two species. Another study confirmed that the broadly used human antibody against γH2AX is applicable in canine cells as well [41]. Although the MVC study is the only investigation of DDR initiation in canine cells, it: (i) implies similarities between human and canine response and (ii) represents an important starting point for exploring the impact of other stressors on canine cells.…”