Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain.
Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain.
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开发改进的方法来鉴定和分析记者小鼠菌株中流式细胞术的肺纤维细胞。
DOI：
10.1002/iid3.361
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Nishioka Y
中科院分区：
文献类型：
--
作者：
Kawano H;Koyama K;Nishimura H;Toyoda Y;Kagawa K;Sato S;Naito N;Goto H;Inagaki Y;Nishioka Y
纤维细胞是新兴的髓源性循环细胞，可以迁移到受损组织中，通常有助于修复。纤维细胞的主要特征包括表达髓样标志物、产生细胞外基质蛋白和分泌激活常驻成纤维细胞的各种体液因子;它们也具有分化成成纤维细胞的潜力。然而，没有特定的表面标记物已被确定为在体内识别纤维细胞。一个原因可能是用于检测纤维细胞的方法需要细胞内胶原染色。在本研究中，为了建立一种用于检测肺纤维细胞并分析活纤维细胞的改进方法，我们使用I型胶原（α）2绿色荧光蛋白（Col‐GFP）报告小鼠，该小鼠经历了博莱霉素（BLM）的肺内滴注。使用流式细胞术门控具有自发荧光的细胞，我们清楚地发现CD 45 + GFP+细胞以稳态驻留在Col‐GFP小鼠的肺中，并且这些细胞在BLM损伤后增加，在第14天达到峰值。这些细胞不仅表达纤维细胞的已知细胞表面标志物，而且还表达一些新的标志物，除了与CD 45 − GFP+细胞相比低水平的胶原蛋白I之外。我们的研究结果表明，改进的方法可以用于检测纯肺纤维细胞，并允许我们进一步分析活纤维细胞的特性。本文建立了一种改良的肺纤维细胞检测方法。使用流式细胞术以自身荧光和联合收割机结合CD 11b、CD 11 c、Gr-1的表达模式来门控细胞，我们清楚地发现纤维细胞驻留在肺中。
Fibrocytes are emerging myeloid‐derived circulating cells that can migrate into damaged tissues and usually contribute to their repair. Key features of fibrocytes include the expression myeloid markers, production of extracellular matrix proteins, and secretion of various humoral factors that activate resident fibroblasts; they also have the potential to differentiate into fibroblasts. However, no specific surface markers have been identified to identify fibrocytes in vivo. One reason could be that the method used to detect fibrocytes requires intracellular collagen staining. In the present study, to establish an improved method for the detection of lung fibrocytes and to analyze viable fibrocytes, we used collagen I(α)2‐green fluorescent protein (Col‐GFP) reporter mice, which had undergone the intratracheal instillation of bleomycin (BLM). Using flow cytometry to gate out cells with autofluorescence, we clearly found that CD45+ GFP+ cells resided in the lungs of Col‐GFP mice at a steady state and these cells increased after BLM injury, peaking at Day 14. These cells expressed not only known cell surface markers of fibrocytes, but also some novel markers, in addition to a low level of collagen I in comparison to CD45− GFP+ cells. Our findings suggest that the improved method can be a useful for the detection of pure lung fibrocytes and allows us to further analyze the characteristics of viable fibrocytes. In this report, we established improved method for detection of lung fibrocytes. Using flow cytometry to gate out cells with autofluorescence and combine expression pattern of CD11b, CD11c, Gr‐1, we clearly found fibrocytes resided in lungs.
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DOI：
10.1038/nri2990
发表时间：
2011-06
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1165/rcmb.2017-0301ps
发表时间：
2018-03-01
影响因子：
6.4
作者：
Dupin, Isabelle;Contin-Bordes, Cecile;Berger, Patrick
通讯作者：
Berger, Patrick
影响因子：
5
作者：
Quan, Timothy E;Cowper, Shawn E;Bucala, Richard
通讯作者：
Bucala, Richard
影响因子：
15.9
作者：
Phillips, RJ;Burdick, MD;Strieter, RM
通讯作者：
Strieter, RM
影响因子：
120.7
作者：
Raghu, Ganesh;van den Blink, Bernt;Richeldi, Luca
通讯作者：
Richeldi, Luca