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用于生物人工肝系统开发的永生化人肝细胞系的建立与鉴定

Establishment and characterization of an immortalized human hepatocyte line for the development of bioartificial liver system.

作者信息

Yan Qun, Deng Lijuan, Zhao Xinmei, Ye Liangying, Fang Yuxin, Meng Yan, Wang Zenan, Luo Xiaobei, Liu Side, Li Aimin

机构信息

Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China.

Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore.

出版信息

Cytotechnology. 2018 Apr;70(2):665-674. doi: 10.1007/s10616-017-0167-3. Epub 2018 Feb 12.

DOI:10.1007/s10616-017-0167-3
PMID:29435697
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5851960/
Abstract

For further research and application of Bioartificial liver systems (BAL), active proliferation capacity and full hepatic functionality of the biomaterials is mandatory. However, there are still no suitable cell lines meeting the requirements for an ideal cell source in BAL development that makes it necessary to explore other sources. Here, we constructed a new cell line derived from well-differentiated hepatocellular carcinoma tissues designated NHBL2. Immunol staining showed that NHBL2 possessed the capacity of synthesizing albumin and CYP2E1 and quantitative analysis showed that the albumin synthesis ability of NHBL2 was comparable to C3A while urea production was highly abundant of NHBL2 compared with that of C3A. Using gene expression microarray analysis, we found that the expression levels of a set of genes encoding Phase I and Phase II metabolizing enzymes as well as many others related to common bioconversion and metabolic processes were significantly higher in NHBL2 cell line than that in C3A. Moreover, functional optimization assay in Matrigel showed obvious improvements of liver-related function level and a low malignance of this cell line. These findings indicated that NHBL2 possessed relatively attractive and full hepatic functionality that might be a potential cell line for BAL development.

摘要

为了生物人工肝系统(BAL)的进一步研究和应用,生物材料的活跃增殖能力和完整的肝脏功能是必不可少的。然而,在BAL开发中,仍然没有合适的细胞系能够满足理想细胞来源的要求,这使得有必要探索其他来源。在此,我们构建了一种源自高分化肝细胞癌组织的新细胞系,命名为NHBL2。免疫组化染色显示NHBL2具有合成白蛋白和CYP2E1的能力,定量分析表明NHBL2的白蛋白合成能力与C3A相当,而与C3A相比,NHBL2的尿素生成量非常丰富。通过基因表达微阵列分析,我们发现NHBL2细胞系中一组编码I相和II相代谢酶以及许多其他与常见生物转化和代谢过程相关的基因的表达水平明显高于C3A。此外,在基质胶中的功能优化试验表明该细胞系的肝脏相关功能水平有明显改善且恶性程度较低。这些发现表明NHBL2具有相对吸引人的完整肝脏功能,可能是一种用于BAL开发的潜在细胞系。