1] Department of Biochemistry and Molecular Pharmacology, Graduate School of Medicine, Chiba University, Chiba, Japan [2] Department of Respirology, Graduate School of Medicine, Chiba University, Chiba, Japan.
Department of Biochemistry and Molecular Pharmacology, Graduate School of Medicine, Chiba University, Chiba, Japan.
Lab Invest. 2014 Nov;94(11):1247-59. doi: 10.1038/labinvest.2014.109. Epub 2014 Sep 8.
Cell-based therapy is recognized as one of potential therapeutic options for lung fibrosis. However, preparing stem/progenitor cells is complicated and not always efficient. Here, we show easily prepared cell populations having therapeutic capacity for lung inflammatory disease that are named as 'lung mixed culture-derived epithelial cells' (LMDECs). LMDECs expressed surfactant protein (SP)-C and gave rise to type I alveolar epithelial cells (AECs) in vitro and in vivo that partly satisfied type II AEC-like characteristics. An intratracheal delivery of not HEK 293 cells but LMDECs to the lung ameliorated bleomycin (BLM)-induced lung injury. A comprehensive analysis of bronchoalveolar fluid by western blot array revealed that LMDEC engraftment could improve the microenvironment in the BLM-instilled lung in association with stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 signaling axis. SDF-1 enhanced both migration activity and differentiating efficiency of LMDECs. Further classification of LMDECs by flow cytometric study showed that a major population of LMDECs (LMDEC(Maj), 84% of total LMDECs) was simultaneously SP-C(+), CD44(+), CD45(+), and hematopoietic cell lineage(+) and that LMDECs included bronchioalveolar stem cells (BASCs) showing SP-C(+)Clara cell secretory protein(+)stem cell antigen (Sca)1(+) as a small population (1.8% of total LMDECs). CD44(+)-sorted LMDEC(Maj) and Sca1(+)-sorted LMDECs equally ameliorated fibrosis induced by BLM like LMDECs did. However, infiltrated neutrophils were observed in Sca1(+)-sorted LMDEC-treated alveoli that was not typical in LMDEC(Maj)- or LMDEC-treated alveoli. These findings suggest that the protective effect of LMDECs against BLM-induced lung injury depends greatly on that of LMDEC(Maj). Furthermore, the cells expressing both alveolar epithelial and hematopoietic cell lineage markers (SP-C(+)CD45(+)) that have characteristics corresponding to LMDEC(Maj) were observed in the alveoli of lung and increased approximately threefold in response to BLM instillation. Taken together, LMDECs newly classified in the present study are easily culture expanded and have a potential role in future regenerative cell therapy for pulmonary fibrosis.
细胞疗法被认为是肺纤维化的潜在治疗选择之一。然而，制备干细胞/祖细胞很复杂，并不总是有效。在这里，我们展示了易于制备的具有治疗肺炎症性疾病能力的细胞群体，这些细胞被称为“肺混合培养衍生的上皮细胞”（LMDECs）。LMDECs 表达表面活性剂蛋白 (SP)-C，并在体外和体内产生 I 型肺泡上皮细胞 (AECs)，部分满足 II 型 AEC 样特征。将 HEK293 细胞而不是 LMDECs 通过气管内给药递送到肺部，可以改善博来霉素（BLM）诱导的肺损伤。通过 Western blot 阵列对支气管肺泡液进行的综合分析表明，LMDEC 移植可以改善 BLM 灌注肺中的微环境，与基质细胞衍生因子-1（SDF-1）/CXC 趋化因子受体 4 信号轴有关。SDF-1 增强了 LMDECs 的迁移活性和分化效率。通过流式细胞术研究对 LMDECs 进行进一步分类表明，LMDECs 的主要群体（LMDEC（Maj），占总 LMDECs 的 84%）同时是 SP-C(+)、CD44(+)、CD45(+)和造血细胞谱系(+)，并且 LMDECs 包括支气管肺泡干细胞（BASCs），表现为 SP-C(+)Clara 细胞分泌蛋白(+)干细胞抗原（Sca）1(+)作为一个小群体（占总 LMDECs 的 1.8%）。CD44(+)分选的 LMDEC（Maj）和 Sca1(+)分选的 LMDECs 与 LMDECs 一样，同样改善了 BLM 诱导的纤维化。然而，在 Sca1(+)分选的 LMDEC 处理的肺泡中观察到浸润的中性粒细胞，这在 LMDEC（Maj）或 LMDEC 处理的肺泡中并不典型。这些发现表明，LMDECs 对 BLM 诱导的肺损伤的保护作用在很大程度上取决于 LMDEC（Maj）。此外，在 BLM 灌注后，肺泡中观察到表达肺泡上皮细胞和造血细胞谱系标志物（SP-C(+)CD45(+)）的细胞，数量增加了约三倍。综上所述，本研究中新分类的 LMDECs 易于培养扩增，在未来肺纤维化的再生细胞治疗中具有潜在作用。
Zotero 插件