外耳道鳞状细胞癌（EACSCC）是一种极其罕见且具有侵袭性的恶性疾病。建立具有强大分子特征的EACSCC细胞系对于EACSCC的基础和转化研究至关重要。在这里，我们展示了新建立的 EACSCC 细胞系 SCEACono2，该细胞系源自患有中分化 EACSCC 的患者。我们利用包括下一代测序 (NGS) 在内的多项实验分析了 SCEACono2 的组织学和遗传特征。 SCEACono2 的免疫细胞化学染色显示 p53 和 SCC1/2 呈阳性。此外，SCEACono2表现出独特的特征，即细胞角蛋白、波形蛋白以及癌症干细胞标志物（CD44、CD133、ALP和Oct3/4）呈阳性。 SCEACono2能够在颞部病变异种移植裸鼠模型中形成肿瘤。 NGS 显示 SCEACono2 含有 TP53 (p.G245S) 和 NOTCH1 (p.A465T) 的体细胞突变。 RNA-seq 和下游生物信息学分析显示，与 SCC-9 和 HSC-4 相比，SCEACono2 中涉及炎症和细胞粘附的基因显着富集。 STR 分析表明没有交叉污染的证据。总之，SCEACono2 可以作为 EACSCC 体外和体内有前途且强大的研究资源。© 2023。作者。

Squamous cell carcinoma of the external auditory canal (EACSCC) is an extraordinarily rare and aggressive malignant disease. Establishment of EACSCC cell line with robust molecular characteristics is essential for the basic and translational research of EACSCC. Here, we show the newly established EACSCC cell line SCEACono2, derived from a patient with well-to-moderately differentiated EACSCC. We analyzed histologic and genetic features of SCEACono2 hiring multiple experiments, including next-generation sequencing (NGS). Immunocytochemical staining of SCEACono2 showed positivity of p53 and SCC1/2. Furthermore, SCEACono2 exhibited a unique characteristic that cytokeratin, vimentin as well as cancer stem cell markers (CD44, CD133, ALP and Oct3/4) were positive. SCEACono2 had an ability to form tumors at the temporal lesion xenograft nude mice model. NGS revealed that SCEACono2 harbored the somatic mutations of TP53 (p.G245S) and NOTCH1 (p.A465T). RNA-seq and downstream bioinformatics analysis revealed significant enrichment of genes involved in inflammation and cell adhesion in SCEACono2 compared to SCC-9 and HSC-4. STR profiling indicated no evidence of cross-contamination. In conclusion, SCEACono2 could serves as a promising and robust research resource of EACSCC in vitro and in vivo.© 2023. The Author(s).