Mimicking the niche of lung epithelial stem cells and characterization of several effectors of their in vitro behavior
Mimicking the niche of lung epithelial stem cells and characterization of several effectors of their in vitro behavior
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DOI：
10.1016/j.scr.2015.05.005
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发表时间：
2015-07-01
影响因子：
1.2
通讯作者：
Betsuyaku, Tomoko
中科院分区：
文献类型：
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作者：
Hegab, Ahmed E.;Arai, Daisuke;Betsuyaku, Tomoko
干细胞周围的小生境在体内平衡期间和损伤或感染后调节它们的命运。3D类器官测定已被广泛用于研究干细胞行为，基于其评估自我更新、分化以及各种培养基补充剂、药物和与支持细胞共培养的效果的能力。我们建立了一种体外研究肺和气管干细胞的方法。我们在基线时表征了它们的增殖和分化谱，然后评估了与成纤维细胞和内皮细胞共培养和/或用几种生物学相关物质处理的效果，作为它们的生态位的可能贡献者。我们发现，肺上皮（而不是气管基底）干细胞需要与基质细胞共培养进行克隆增殖和分化。成纤维细胞在提供这种支持方面比内皮细胞更有效，并且支持模式根据基质细胞的组织来源而变化。用FGF 2、FGF 9、FGF 10、LIF以及ALK 5和ROCK抑制剂处理远端肺上皮或基底干细胞增加了它们的集落形成效率，并对集落数量、大小和分化谱产生了不同的影响。这个模型和发现为更好地理解肺干细胞生态位成分和可以操纵肺干细胞行为的因素铺平了道路。(C)2015作者出版社：Elsevier B. V.
The niche surrounding stem cells regulate their fate during homeostasis and after injury or infection. The 3D organoid assay has been widely used to study stem cells behavior based on its capacity to evaluate self-renewal, differentiation and the effect of various medium supplements, drugs and co-culture with supportive cells. We established an assay to study both lung and trachea stem cells in vitro. We characterized their proliferation and differentiation spectrum at baseline then evaluated the effect of co-culturing with fibroblasts and endothelial cells and/or treating with several biologically relevant substances as possible contributors to their niche. We found that lung epithelial (but not tracheal basal) stem cells require co-culture with stromal cells to undergo clonal proliferation and differentiation. Fibroblasts were more efficient than endothelial cells in offering this support and the pattern of support varied based on the tissue origin of the stromal cells. Treating distal lung epithelial or basal stem cells with FGF2, FGF9, FGF10, LIF as well as ALK5 and ROCK inhibitors increased their colony formation efficiency and resulted in variable effects on colonies number, size and differentiation spectrum. This model and findings pave the way for better understanding of lung stem cell niche components and factors that can manipulate lung stem cell behavior. (C) 2015 The Authors. Published by Elsevier B.V.