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全面的基因和分子特征分析证实了永生化Col-GFP肝星状细胞系的肝星状细胞起源。

Comprehensive Genetic and Molecular Characterization Confirms Hepatic Stellate Cell Origin of the Immortal Col-GFP HSC Line.

作者信息

Buitkamp Larissa F, Liehr Thomas, Kankel Stefanie, Buhl Eva M, Hardt Katharina S, Keller Diandra T, Schröder-Lange Sarah K, Weiskirchen Ralf

机构信息

Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry (IFMPEGKC), RWTH University Hospital Aachen, D-52074 Aachen, Germany.

Jena University Hospital, Institute of Human Genetics, Friedrich Schiller University, D-07747 Jena, Germany.

出版信息

Int J Mol Sci. 2025 Aug 11;26(16):7764. doi: 10.3390/ijms26167764.

Abstract

The immortal murine hepatic stellate cell line Col-GFP HSC was comprehensively characterized using genetic and molecular approaches. Short tandem repeat (STR) profiling and karyotyping combined with multiplex fluorescence in situ hybridization (M-FISH) confirmed the identity of the cell line and revealed no contamination. Col-GFP HSCs showed a near tetraploid karyotype. Additionally, next-generation sequencing (NGS) data, quantitative reverse transcription PCR, and Western blot analyses demonstrated robust expression of genes and proteins associated with hepatic stellate cells, including those involved in extracellular matrix remodeling and fibrogenic pathways. Phalloidin staining revealed filamentous actin patterns characteristic of stellate cells, providing additional support for their cytoskeletal organization and functional status. These findings provide strong evidence that the Col-GFP HSC cell line originates from hepatic stellate cells and can serve as a reliable in vitro model to study stellate cell biology and related pathophysiological processes.

摘要

使用遗传和分子方法对永生化小鼠肝星状细胞系Col-GFP HSC进行了全面表征。短串联重复序列(STR)分析和核型分析结合多重荧光原位杂交(M-FISH)证实了该细胞系的身份,且未发现污染。Col-GFP HSCs显示出近四倍体核型。此外,下一代测序(NGS)数据、定量逆转录PCR和蛋白质印迹分析表明,与肝星状细胞相关的基因和蛋白质表达强劲,包括那些参与细胞外基质重塑和纤维化途径的基因和蛋白质。鬼笔环肽染色揭示了星状细胞特有的丝状肌动蛋白模式,为其细胞骨架组织和功能状态提供了额外支持。这些发现提供了强有力的证据,表明Col-GFP HSC细胞系源自肝星状细胞,可作为研究星状细胞生物学和相关病理生理过程的可靠体外模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3909/12386721/84fc6d7a1e3a/ijms-26-07764-g001.jpg

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