Clinical Research Center, The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210003, P.R. China.
Department of Pathology, The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing 210003, P.R. China.
Exp Biol Med (Maywood). 2022 Jun;247(11):972-981. doi: 10.1177/15353702221085534. Epub 2022 Apr 26.
Liver cancer (LC) is one of the most common malignant tumors worldwide. Since the mechanism of LC pathogenesis and metastasis cannot be carried out directly on the human body, it is particularly important to establish human liver cancer cell lines for research . In this study, tissue block adherence method combined with cell clumps digestion method was used to establish primary human hepatocytes (PHHs) with a successful rate of 60% (45/75). Short tandem repeat (STR) analysis proved the cells were derived from its paired tissues. These cells from hepatocellular carcinoma (HCC) expressed NTCP and secreted ALB and AAT as detected by western blot, and expressed hepatocyte-specific membrane protein ASGR1 as detected by flow cytometry. Liver cancer biomarkers like CK7 in ICC (intrahepatic cholangiocarcinoma), AFP, and GPC3 in HCC expressed of different degree as detected by immunohistochemical analysis. These cells displayed typical liver cancer cell morphological characteristics and can passage stably. In conclusion, we developed an effective method to establish PHHs. Further studies are necessary to study if these cells maintaining other liver function and reproduce the physiology of the tumors and how these cells behavior in the drug development.
肝癌 (LC) 是全球最常见的恶性肿瘤之一。由于不能直接在人体上进行 LC 发病机制和转移的研究，因此建立人肝癌细胞系对于研究非常重要。在本研究中，我们采用组织块贴壁法联合细胞团消化法，成功建立了原代人肝癌细胞 (PHH)，成功率为 60%（45/75）。短串联重复序列 (STR) 分析证明这些细胞来源于其配对组织。Western blot 检测表明，这些细胞表达 NTCP 并分泌 ALB 和 AAT，流式细胞术检测表明细胞表达肝特异性膜蛋白 ASGR1。免疫组织化学分析显示 ICC（肝内胆管癌）中的 CK7、HCC 中的 AFP 和 GPC3 等肝癌标志物的表达程度不同。这些细胞呈现出典型的肝癌细胞形态特征，并能稳定传代。总之，我们开发了一种有效的建立 PHH 的方法。进一步的研究需要研究这些细胞是否能维持其他肝脏功能，重现肿瘤的生理学特性，以及这些细胞在药物开发中的行为方式。
