Human hepatic stellate cell lines, LX-1 and LX-2: new tools for analysis of hepatic fibrosis
Human hepatic stellate cell lines, LX-1 and LX-2: new tools for analysis of hepatic fibrosis
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DOI：
10.1136/gut.2004.042127
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发表时间：
2005-01-01
期刊：
影响因子：
24.5
通讯作者：
Eng, FJ
中科院分区：
文献类型：
--
作者：
Xu, L;Hui, AY;Eng, FJ
背景：肝星状细胞（HSC）是一种主要的纤维化细胞类型，在慢性肝病期间有助于胶原积累。随着开发抗纤维化疗法的兴趣日益增加，需要保留人HSC的体内表型的细胞系来阐明人肝纤维化的途径。我们建立了两个人HSC细胞系LX-1和LX-2，并将其特征与原代人stellate cells.Methods和Results进行了比较：LX-1和LX-2是由SV 40 T抗原永生化（LX-1）或低血清条件下自发永生化（LX-2）产生的。这两条线表达平滑肌肌动蛋白，波形蛋白和胶质细胞酸性蛋白，如通过免疫细胞化学观察到的。与原代HSC类似，两种细胞系均表达调节肝纤维化的关键受体，包括血小板衍生生长因子受体β（β PDGF-R）、肥胖受体长型（Ob-R-L）和盘状结构域受体2（DDR2），以及参与基质重塑的蛋白质;基质金属蛋白酶（MMP）-2、基质金属蛋白酶组织抑制剂（TIMP）-2和MT 1-MMP，如通过Western分析确定的。LX-2的TIMP-1表达降低。LX-2，而不是LX-1，响应于PDGF而增殖。两种细胞系均表达α 1（I）前胶原和HSP 47的mRNA。转化生长因子β 1刺激增加他们的α 1（I）前胶原mRNA的表达，通过定量逆转录聚合酶链反应测定。LX-2细胞，而不是LX-1细胞，是高度可转染的。这两种细胞系都具有典型的星状细胞维甲酸表型。基因芯片分析表明，原代HSC与LX-1（98.4%）或LX-2（98.7%）之间的基因表达具有很强的相似性，具有多个神经元基因的表达。结论：LX-1和LX-2人HSC系在肝脏疾病的研究提供了有价值的新工具。这两种细胞系都保留了HSC的关键特征。LX-2的两个独特优势是其在无血清培养基中的活力和高转染性。
Background: Hepatic stellate cells (HSCs) are a major fibrogenic cell type that contributes to collagen accumulation during chronic liver disease. With increasing interest in developing antifibrotic therapies, there is a need for cell lines that preserve the in vivo phenotype of human HSCs to elucidate pathways of human hepatic fibrosis. We established and characterised two human HSC cell lines termed LX-1 and LX-2, and compared their features with those of primary human stellate cells.Methods and results: LX-1 and LX-2 were generated by either SV40 T antigen immortalisation (LX-1) or spontaneous immortalisation in low serum conditions (LX-2). Both lines express a smooth muscle actin, vimentin, and glial fibrillary acid protein, as visualised by immunocytochemistry. Similar to primary HSCs, both lines express key receptors regulating hepatic fibrosis, including platelet derived growth factor receptor beta (betaPDGF-R), obese receptor long form (Ob-R-L), and discoidin domain receptor 2 (DDR2), and also proteins involved in matrix remodelling; matrix metalloproteinase (MMP)-2, tissue inhibitor of matrix metalloproteinase (TIMP)-2, and MT1-MMP, as determined by western analyses. LX-2 have reduced expression of TIMP-1. LX-2, but not LX-1, proliferate in response to PDGF. Both lines express mRNAs for alpha1(I) procollagen and HSP47. Transforming growth factor beta1 stimulation increased their alpha1(I) procollagen mRNA expression, as determined by quantitative reverse transcription-polymerase chain reaction. LX-2, but not LX-1, cells are highly transfectable. Both lines had a retinoid phenotype typical of stellate cells. Microarray analyses showed strong similarity in gene expression between primary HSCs and either LX-1 (98.4%) or LX-2 (98.7%), with expression of multiple neuronal genes.Conclusions: LX-1 and LX-2 human HSC lines provide valuable new tools in the study of liver disease. Both lines retain key features of HSCs. Two unique advantages of LX-2 are their viability in serum free media and high transfectability.