Detection of abundant megakaryocytes in pulmonary artery blood in lung cancer patients using a microfluidic platform
Detection of abundant megakaryocytes in pulmonary artery blood in lung cancer patients using a microfluidic platform
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DOI：
10.1016/j.lungcan.2018.09.011
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发表时间：
2018-11-01
期刊：
影响因子：
5.3
通讯作者：
Sakao, Yukinori
中科院分区：
文献类型：
--
作者：
Dejima, Hitoshi;Nakanishi, Hayao;Sakao, Yukinori
目的：最近在小鼠体内重新发现肺是一种生成血小板的造血器官。然而，肺在人类血栓形成中的作用的证据仍然有限。在这项研究中，我们利用一个新开发的稀有细胞分离微流控平台，检测了肺和体循环中的巨核细胞，特别是肺动脉血(PAB)、静脉血(PVB)和外周血中的巨核细胞。材料和方法：我们分析了23例在我所接受手术的肺癌患者。术后即刻取肺组织行PAB和PVB。使用基于过滤的微流控装置对血液样本进行大小选择，并对玻片上浓缩的稀有细胞进行巴氏染色(Pap)、免疫细胞化学(ICC)和免疫荧光(IF)。结果：PAP/ICC/IF显示PAB中有丰富的CD61+/细胞角蛋白巨细胞，并有巨核细胞谱系，而PVB中仅有少量CD61+/细胞角蛋白巨细胞。这些巨核细胞由CD61+/CD41+未成熟巨核细胞和CD61+/CD41-成熟巨核细胞组成，具有产生血小板的能力。这些发现得到了Giemsa染色的血涂片常规血液学分析的证实。在肺癌分析中，CD61+巨核细胞仅见于非癌组织的毛细血管中，而血小板则选择性地见于癌组织的肿瘤血管中。结论：这些结果表明，大量的巨核细胞从全身骨髓迁移到PAS中聚集，成熟的巨核细胞在正常肺的毛细血管中滞留，提示肺在肺癌患者全身的血小板生成中可能起着生理作用。
Objectives: The lung was recently re-discovered as a hematopoietic organ for platelet production in mice. However, evidence for the role of the lung in thrombopoiesis in humans is still limited. In this study, we examined megakaryocytes in the pulmonary and systemic circulation, specifically in pulmonary arterial blood (PAB), venous blood (PVB) and peripheral blood using a newly developed microfluidic platform for rare cell isolation.Materials and methods: We analyzed 23 lung cancer patients who underwent surgery in our institute. PAB and PVB were obtained from the resected lung immediately after surgery. Blood samples were size-selected using a filtration-based microfluidic device and enriched rare cells on glass slide specimens were stained with Papanicolaou (Pap), immunocytochemistry (ICC), and immunofluorescence (IF). Lung tissues were also analyzed by immunohistochemistry.Results: Pap/ICC/IF showed the presence of abundant CD61 + /cytokeratin- giant cells with a megakaryocyte lineage in PAB, but only a few in PVB. These megakaryocytes were found to consist of CD61 + /CD41 + immature megakaryocytes and CD61 +/CD41- mature megakaryocytes with the potential to produce platelets. These findings were confirmed by the conventional hematological analysis of blood smears stained with Giemsa. In analysis of lung cancer, CD61 + megakaryocytes were observed exclusively in the capillaries of non-cancerous tissue, whereas platelets were selectively observed in the tumor blood vessels of cancerous tissue.Conclusions: These results indicate that numerous megakaryocytes migrate from systemic bone marrows to accumulate in PAs and arrest of mature megakaryocytes in the capillaries of normal lung, suggesting the possibility that the lung plays a physiological role in the systemic thrombopoiesis in lung cancer patients.