Direct analysis of hepatic stellate cells with flow cytometry in specimens derived from the human liver.

IF 3.2 4区医学 Q2 IMMUNOLOGY

International immunology Pub Date : 2025-09-15 DOI:10.1093/intimm/dxaf055

Toshiaki Bando, Hirotaka Sato, Shunsuke Uno, Hajime Morita, Lynn Zreka, Shuhe Ma, Mouna Khan, Daichi Akuzawa, Yuki Masuo, Joey Matsuyama, Ryo Nishida, Shinya Okumura, Etsuro Hatano, Takashi Ito, Hideki Ueno

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引用次数: 0

Abstract

Background & aims: Hepatic stellate cells (HSCs) play a crucial role in liver fibrosis. However, the methodology to directly assess the biology of primary HSCs in human liver specimens is yet to be established. In this study, we aimed to establish a robust methodology to analyze primary HSCs in human liver specimens with flow cytometry (FCM).

Methods: We first applied FCM to HSCs directly isolated from liver tissues with Nycodenz density gradients. Then, we analyzed HSCs in frozen/thawed liver perfusate samples and liver tissues. We also compared the phenotype of HSCs in primary biliary cholangitis (PBC) and those in healthy counterparts.

Results: We found that HSCs were substantially smaller and less dense than normal lymphocytes in the FCM analysis. By carefully defining the FCM gating strategy, we were able to establish the approach to analyze both quiescent HSCs (qHSCs) and activated HSCs (aHSCs) in human liver specimens. Importantly, we found that co-expression of CD14 and CD56 within CD45neg non-immune cells permits the detection of qHSCs, whereas CD68 and CD40 within CD45neg non-immune cells were valuable for assessing aHSCs. Furthermore, we found that aHSCs in PBC upregulated the expression of multiple markers associated with antigen-presentation capacity.

Conclusion: Our established approach with FCM will be valuable for the direct analysis of qHSCs and aHSCs with FCM in various human liver specimens. Our FCM analysis of aHSCs in PBC suggested their involvement in the local immune responses.

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用流式细胞术直接分析人肝脏标本中的肝星状细胞。

背景与目的：肝星状细胞（HSCs）在肝纤维化中起重要作用。然而，直接评估人肝脏标本中原代造血干细胞生物学的方法尚未建立。在这项研究中，我们旨在建立一种强大的方法，用流式细胞术（FCM）分析人肝脏标本中的原代造血干细胞。方法：首先采用流式细胞术对具有Nycodenz密度梯度的肝组织直接分离的造血干细胞进行培养。然后，我们分析了冷冻/解冻肝灌注液样本和肝组织中的造血干细胞。我们还比较了原发性胆道胆管炎（PBC）患者和健康人群中造血干细胞的表型。结果：在流式细胞术中，我们发现造血干细胞比正常淋巴细胞体积更小，密度更低。通过仔细定义FCM门控策略，我们能够建立分析人肝脏标本中静止hsc （qhsc）和激活hsc （aHSCs）的方法。重要的是，我们发现CD14和CD56在cd45阴性非免疫细胞内的共表达允许检测qhsc，而CD68和CD40在cd45阴性非免疫细胞内对评估aHSCs有价值。此外，我们发现PBC中的aHSCs上调了与抗原呈递能力相关的多种标志物的表达。结论：本研究建立的FCM方法可用于多种人肝脏标本qhsc和aHSCs的FCM直接分析。我们对PBC中aHSCs的流式细胞仪分析表明它们参与了局部免疫反应。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International immunology 医学-免疫学

CiteScore

9.30

自引率

2.30%

发文量

审稿时长

6-12 weeks

期刊介绍： International Immunology is an online only (from Jan 2018) journal that publishes basic research and clinical studies from all areas of immunology and includes research conducted in laboratories throughout the world.