In the global progress of bone tumor research, established stable and long-lasting transgenic chondrosarcoma (CSA) cell lines are rare, mainly of murine and human origin, while the establishment of canine CSA cell lines has yet to be reported. This study established a canine CSA cell line to facilitate the basic clinical study of canine CSA. Fifty five cases of canine osteolytic disease were collected, and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture. A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully cultured. According to the clinical characteristics of the dog and the histopathological results of the primary tumor, CSA was diagnosed, and the CSA cell line was designated Mango. Immunohistochemical (IHC) results showed that the immunoreactivity of bone gamma-carboxyglutamate protein (BGLAP), secreted protein acidic and rich in cysteine (SPARC), alkaline phosphatase (ALPL), vimentin (VIM) and S100 were positive. However, the immunoreactivity of pan-cytokeratin (PCK), chromogranin A (CGA), and platelet endothelial cell adhesion molecule-1 (CD31) was negative. Immunofluorescence (IF) results showed that the protein expressions in the Mango cell line were consistent with the IHC identification of the primary tumor. The Mango cell line’s doubling time was 43.92 h, and the cell formation rate exceeded 20%. There were abnormal chromosome numbers, hetero staining with toluidine blue, and certain calcification abilities. It could be passaged stably and continuously without changing the cell morphology and characteristics. In vivo, the cells were successfully injected into the nude mice model with a tumorigenic rate of 100%. The immunophenotype of the xenograft tumor was consistent with that of the primary tumor. Therefore, we effectively established a canine CSA cell line. As a promising cell material, this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic research of canine CSA. Moreover, because of its similarity to human CSA, the animal model of CSA is also indispensable for investigating human CSA.

在全球骨肿瘤研究进展中，建立的稳定且持久的转基因软骨肉瘤（CSA）细胞系很少见，主要来源于小鼠和人类，而犬CSA细胞系的建立尚未见报道。本研究建立了犬CSA细胞系，以促进犬CSA的基础临床研究。收集55例犬溶骨性疾病病例，选取10余例具有典型临床症状的犬骨肿瘤样本进行原代细胞培养。成功培养出稳定传代超过100代且具有小鼠致瘤能力的细胞系。根据狗的临床特征和原发肿瘤的组织病理学结果，诊断为CSA，CSA细胞系命名为Mango。免疫组织化学（IHC）结果显示骨γ-羧基谷氨酸蛋白（BGLAP）、富含半胱氨酸的酸性分泌蛋白（SPARC）、碱性磷酸酶（ALPL）、波形蛋白（VIM）和S100免疫反应性呈阳性。然而，全细胞角蛋白（PCK）、嗜铬粒蛋白A（CGA）和血小板内皮细胞粘附分子-1（CD31）的免疫反应性呈阴性。免疫荧光（IF）结果显示Mango细胞系中蛋白表达与原发肿瘤的IHC鉴定一致。 Mango细胞系倍增时间为43.92 h，细胞形成率超过20%。染色体数目异常，甲苯胺蓝异染色，有一定的钙化能力。可稳定、连续传代，不改变细胞形态和特性。在体内，该细胞成功注射入裸鼠模型，成瘤率达100%。异种移植肿瘤的免疫表型与原发肿瘤一致。因此，我们有效地建立了犬CSA细胞系。该细胞系作为一种很有前景的细胞材料，可用于构建荷瘤模型，有利于犬CSA的后续基础研究。此外，由于其与人类CSA的相似性，CSA动物模型对于研究人类CSA也是必不可少的。