原发性癌细胞反映了肿瘤的遗传背景和表型。具有较高增殖活性的永生化细胞比原代细胞具有优势。该研究的目的是使用携带质粒的人端粒酶逆转录酶（hTERT）基因使原发性卵巢癌（OvCa）细胞永生化，并将其与原代细胞的表型和生物活性进行比较。从两名高级别浆液性卵巢癌患者的腹水中分离出原代 OvCa3 A 和 OvCa7 A 细胞，并使用免疫细胞化学方法、流式细胞术、实时 RT-PCR、Western blot、代谢活性和迁移潜力进行表征。两种永生化卵巢癌细胞系都反映了原发癌细胞的表型，尽管有所修改。 OvCa3 A hTERT 细胞保持 CD73/CD90/CD105 阳性的间充质干细胞表型，CD133 阴性，而 OvCa7 A hTERT 细胞群失去 CD73 表达，但几乎 90% 的细胞表达 CSC 特征的 CD133表型。永生化 OvCa 细胞的 Sox2 和 Oct4 基因表达水平存在差异，这与干性特性相关。 OvCa7 A hTERT 细胞比相应的原代 OvCa 细胞表现出更高的代谢和迁移活性以及 ALDH1 表达。原代细胞系和永生化细胞系均能够形成球状体。新建立的独特永生化细胞系OvCa7 A hTERT，具有浆液性卵巢癌恶性特征，并且具有p53、Pax8的积累以及CD133和CD44分子的过度表达，可能是治疗研究的有用工具。方法，特别是那些针对卵巢癌和临床前 2D 和 3D 模型中的 CSC 的方法。

Primary cancer cells reflect the genetic background and phenotype of a tumor. Immortalized cells with higher proliferation activity have an advantage over primary cells. The aim of the study was to immortalize the primary ovarian cancer (OvCa) cells using the plasmid-carrying human telomerase reverse transcriptase (hTERT) gene and compare their phenotype and biological activity with the primary cells. The primary OvCa3 A and OvCa7 A cells were isolated from the ascitic fluid of two high-grade serous ovarian cancer patients and were characterized using immunocytochemical methods, flow cytometry, real-time RT-PCR, Western blot, metabolic activity, and migratory potential. Both immortalized ovarian cancer cell lines mirrored the phenotype of primary cancer cells, albeit with modifications. The OvCa3 A hTERT cells kept the mesenchymal stem cell phenotype of CD73/CD90/CD105-positivity and were CD133-negative, whereas the cell population of OvCa7 A hTERT lost CD73 expression, but almost 90% of cells expressed the CD133 characteristic for the CSCs phenotype. Immortalized OvCa cells differed in gene expression level with respect to Sox2 and Oct4, which was associated with stemness properties. The OvCa7 A hTERT cells showed higher metabolic and migratory activity and ALDH1 expression than the corresponding primary OvCa cells. Both primary and immortalized cell lines were able to form spheroids. The newly established unique immortalized cell line OvCa7 A hTERT, with the characteristic of a serous ovarian cancer malignancy feature, and with the accumulation of the p53, Pax8, and overexpression of the CD133 and CD44 molecules, may be a useful tool for research on therapeutic approaches, especially those targeting CSCs in ovarian cancer and in preclinical 2D and 3D models.