Department of Otorhinolaryngology-Head and Neck Surgery, CHA Bundang Medical Center, CHA University School of Medicine, Seongnam, Gyeonggi-do, 13496, Republic of Korea.
Western Seoul Center, Korea Basic Science Institute, Seoul, Republic of Korea.
J Cancer Res Clin Oncol. 2020 Oct;146(10):2497-2507. doi: 10.1007/s00432-020-03306-7. Epub 2020 Jul 3.
Tumor explant culture systems can mimic the in vivo tumor microenvironment, proposing as a substitute for preclinical studies for prediction of individual treatment response. Therefore, our study evaluated the potential usefulness of ex vivo tumor explants culture assembled into the cell sheets by anticancer drug screening in patients with head and neck squamous cell carcinoma (HNSCC).
Our model included tumor explants incorporated into cell sheet composing of epithelium and subepithelial stroma using tumor and mucosal samples obtained from the HNSCC patients who underwent surgery. Cell growth, viability, and hypoxia were measured by cell counting kit-8, live/dead assay, propidium iodide, and LOX-1 staining, and were compared among the different treatment groups with vehicle, cisplatin or docetaxel.
Tumor explants stably survived in the cell sheet over 10 days after explantation, whereas most of the explants in non-matrix culture became nonviable within 5-8 days with the significant daily decrease of viability. The live tissue areas of tumor explants in the cell sheet maintained over 30 days without significant changes although hypoxic cell areas gradually increased up to 5 days. Tissue viability and live cancer tissue areas significantly decreased after the treatment of cisplatin or docetaxel in the dose and time-dependent manners.
Our cell sheet-based tumor explants model might be applied to the reliable ex vivo screening for anticancer chemotherapeutics for HNSCC.
肿瘤离体培养系统可以模拟体内肿瘤微环境,可作为临床前研究的替代方法,用于预测个体的治疗反应。因此,我们研究评估了将头颈部鳞状细胞癌(HNSCC)患者的肿瘤离体培养物组装成细胞片,并通过抗癌药物筛选进行体外药敏试验的潜在用途。
我们的模型包括使用从接受手术的 HNSCC 患者获得的肿瘤和粘膜样本,将肿瘤离体培养物纳入由上皮和上皮下基质组成的细胞片中。通过细胞计数试剂盒-8、活/死检测、碘化丙啶和 LOX-1 染色来测量细胞生长、活力和缺氧情况,并在不同的治疗组(溶剂、顺铂或多西他赛)之间进行比较。
肿瘤离体培养物在离体后 10 天内稳定地存活在细胞片中,而在无基质培养中的大多数离体培养物在 5-8 天内失去活力,且活力每天显著下降。尽管缺氧细胞区域在 5 天内逐渐增加,但细胞片中原位组织的活组织区域在 30 天以上保持不变。顺铂或多西他赛在剂量和时间依赖性方式下进行处理后,组织活力和活癌细胞组织区域显著下降。
我们基于细胞片的肿瘤离体培养物模型可用于可靠的体外筛选针对 HNSCC 的抗癌化疗药物。