UNASSIGNED: Effusion samples were collected from three dogs with histologically diagnosed pericardial mesothelioma and used for primary culture. Cultured cells were characterized by immunostaining for pan-cytokeratin AE1/AE3, vimentin, Wilms\' tumor suppressor gene 1 (WT1), and cytokeratin 5 (CK5). To assess the tumorigenic properties of cells in the effusion and generate a xenograft model, the cell suspension was injected into a severe combined immunodeficient (SCID) mouse either subcutaneously (SC) or intraperitoneally (IP). Lastly, chemosensitivity of established primary cultures against four drugs, doxorubicin, vinorelbine, carboplatin, and gemcitabine, by single-agent treatment as well as combination treatment of carboplatin at a fixed concentration, either 10 or 100 μM, and gemcitabine at different concentrations ranging from 0-1000 μM was assessed by cell viability assay.
UNASSIGNED: Primary cultures were successfully generated and characterized by dual positivity for AE1/AE3 and vimentin and positive staining for WT-1 and CK5, confirming the mesothelial origin of the cells. In the xenograft models, SC mouse developed a subcutaneous mass, whereas IP mouse developed multiple intraperitoneal nodules. The masses were histopathologically consistent with mesotheliomas. The chemosensitivity assay revealed that carboplatin had the highest anti-tumor effects among the four tested single-agent treatments. Furthermore, carboplatin at 100 μM combined with gemcitabine at clinically relevant doses demonstrated the augmented anti-tumor effects compared to single-agent treatment.
UNASSIGNED: Primary cultures and xenograft models generated in this study could be useful tools for in vitro and in vivo studies of canine mesothelioma. Carboplatin is a highly effective chemotherapeutic agent against canine mesothelioma when used as a sole agent and in combination with gemcitabine.
■从三只经组织学诊断为心包间皮瘤的狗收集积液样品并用于原代培养。培养的细胞通过全细胞角蛋白AE1/AE3,波形蛋白的免疫染色来表征,威尔姆斯抑癌基因1(WT1),和细胞角蛋白5(CK5)。为了评估积液中细胞的致瘤特性并生成异种移植模型,将细胞悬液皮下(SC)或腹膜内(IP)注射到严重的联合免疫缺陷(SCID)小鼠中。最后,已建立的原代培养物对四种药物的化学敏感性,阿霉素,长春瑞滨,卡铂,和吉西他滨,通过单一药物治疗以及以固定浓度联合治疗卡铂,10或100μM,通过细胞活力测定评估0-1000μM范围内的不同浓度的吉西他滨。
■原代培养物被成功地产生并且通过AE1/AE3和波形蛋白的双重阳性以及WT-1和CK5的阳性染色来表征,从而证实细胞的间皮起源。在异种移植模型中,SC小鼠出现皮下肿块,而IP小鼠出现多个腹膜内结节。肿块在组织病理学上与间皮瘤一致。化学敏感性测定显示,卡铂在四种测试的单药治疗中具有最高的抗肿瘤作用。此外,与单药治疗相比,100μM卡铂联合临床相关剂量的吉西他滨显示出增强的抗肿瘤作用。
■本研究中产生的原代培养物和异种移植模型可能是犬间皮瘤的体外和体内研究的有用工具。当用作唯一药剂并与吉西他滨组合时,卡铂是对抗犬间皮瘤的高效化学治疗剂。