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哮喘和非哮喘供体来源的人端粒酶逆转录酶永生化的人气道平滑肌细胞的表型和功能特征。

Phenotype and Functional Features of Human Telomerase Reverse Transcriptase Immortalized Human Airway Smooth Muscle Cells from Asthmatic and Non-Asthmatic Donors.

机构信息

University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, GRIAC (Groningen Research Institute for Asthma and COPD), Groningen, The Netherlands.

University of Groningen, University Medical Center Groningen, KOLFF Institute, Groningen, The Netherlands.

出版信息

Sci Rep. 2018 Jan 16;8(1):805. doi: 10.1038/s41598-017-18429-0.

Abstract

Asthma is an obstructive respiratory disease characterised by chronic inflammation with airway hyperresponsiveness. In asthmatic airways, there is an increase in airway smooth muscle (ASM) cell bulk, which differs from non-asthmatic ASM in characteristics. This study aimed to assess the usefulness of hTERT immortalisation of human ASM cells as a research tool. Specifically we compared proliferative capacity, inflammatory mediator release and extracellular matrix (ECM) production in hTERT immortalised and parent primary ASM cells from asthmatic and non-asthmatic donors. Our studies revealed no significant differences in proliferation, IL-6 and eotaxin-1 production, or CTGF synthesis between donor-matched parent and hTERT immortalised ASM cell lines. However, deposition of ECM proteins fibronectin and fibulin-1 was significantly lower in immortalised ASM cells compared to corresponding primary cells. Notably, previously reported differences in proliferation and inflammatory mediator release between asthmatic and non-asthmatic ASM cells were retained, but excessive ECM protein deposition in asthmatic ASM cells was lost in hTERT ASM cells. This study shows that hTERT immortalised ASM cells mirror primary ASM cells in proliferation and inflammatory profile characteristics. Moreover, we demonstrate both strengths and weaknesses of this immortalised cell model as a representation of primary ASM cells for future asthma pathophysiological research.

摘要

哮喘是一种阻塞性呼吸系统疾病,其特征是慢性炎症和气道高反应性。在哮喘气道中,气道平滑肌(ASM)细胞体积增加,这与非哮喘 ASM 细胞在特性上有所不同。本研究旨在评估 hTERT 永生化人 ASM 细胞作为研究工具的有用性。具体来说,我们比较了哮喘和非哮喘供体的 hTERT 永生化和原代 ASM 细胞的增殖能力、炎症介质释放和细胞外基质(ECM)产生。我们的研究表明,在供体匹配的亲本和 hTERT 永生化 ASM 细胞系之间,增殖、IL-6 和嗜酸性粒细胞趋化因子-1 产生或 CTGF 合成没有显著差异。然而,与相应的原代细胞相比,永生化 ASM 细胞中 ECM 蛋白纤维连接蛋白和纤维蛋白素-1 的沉积显著降低。值得注意的是,先前报道的哮喘和非哮喘 ASM 细胞之间在增殖和炎症介质释放方面的差异得以保留,但在 hTERT ASM 细胞中,哮喘 ASM 细胞中过多的 ECM 蛋白沉积消失了。本研究表明,hTERT 永生化 ASM 细胞在增殖和炎症特征方面与原代 ASM 细胞相似。此外,我们展示了这种永生化细胞模型作为未来哮喘病理生理学研究中代表原代 ASM 细胞的优势和劣势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d20/5770384/94b85a6db84e/41598_2017_18429_Fig1_HTML.jpg

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