Human Tracheobronchial Basal Cells Normal versus Remodeling/Repairing Phenotypes In Vivo and In Vitro
Human Tracheobronchial Basal Cells Normal versus Remodeling/Repairing Phenotypes In Vivo and In Vitro
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DOI：
10.1165/rcmb.2013-0049oc
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发表时间：
2013-12-01
影响因子：
6.4
通讯作者：
Reynolds, Susan D.
中科院分区：
文献类型：
--
作者：
Ghosh, Moumita;Ahmad, Shama;Reynolds, Susan D.
人气管支气管上皮（TBE）基底细胞（BC）在正常组织中起着祖细胞的作用。然而，机制研究通常在体外进行，并且经常使用从死于非呼吸系统疾病的患者中回收的BC。目前尚不清楚尸体上皮是否（1）正在经历稳态重塑和/或修复，或（2）产生代表组织中鉴定的稳态过程的BC克隆。我们试图比较TBE-BCs的表型与在最佳克隆形成条件下培养的BCs的表型。采用定量组织形态计量学评价TBE病理学。通过荧光激活细胞分选仪分析确定培养的BC表型。克隆组织和细胞表型通过免疫染色确定。尸体上的总BE正常20%。在这些区域中，BC是角蛋白（K）-5（+）和四跨膜蛋白CD 151（+），并且显示出低的有丝分裂指数。相比之下，80%的尸体TBE表现出稳态重塑/修复过程。在这些区域中，BC是K5（+）/K14（+），并且一个子集表达组织因子（TF）。第1代TBE细胞是K5（+）/TF+的BC，并且半共表达CD 151。最佳克隆形成条件使用经辐照的NIH 3 T3成纤维细胞饲养层（美国典型培养物保藏中心，Frederick，MD）和补充血清的Epicult-B培养基（Stem cell Technologies，拉霍亚，CA）。TF+/CD 151（-）BC亚群是最具克隆形成性的BC亚型，并且富含K14（+）细胞。TF+/CD 151（-）BC产生含有K5（+）/Trp 63（+）但K14（-）/CD 151（-）BC的克隆。TF+细胞仅限于克隆边缘。总之，克隆原性人TBE BC（1）表现出一种分子表型，其是在组织中观察到的正常和重塑/修复性BC表型的复合物，并且（2）产生含有表型不同的BC亚群的类器官克隆。
Human tracheobronchial epithelial (TBE) basal cells (BCs) function as progenitors in normal tissue. However, mechanistic studies are typically performed in vitro and frequently use BCs recovered from patients who die of non respiratory disease. It is not known whether the cadaveric epithelium (1) is undergoing homeostatic remodeling and/or repair, or (2) yields BC clones that represent homeostatic processes identified in tissue. We sought to compare the phenotype of TBE-BCs with that of BCs cultured under optimal clone-forming conditions. TBE pathology was evaluated using quantitative histomorphometry. The cultured BC phenotype was determined by fluorescence-activated cell sorter analysis. Clone organization and cell phenotype were determined by immunostaining. The cadaveric TBE is 20% normal. In these regions, BCs are keratin (K)-5(+) and tetraspanin CD151(+), and demonstrate a low mitotic index. In contrast, 80% of the cadaveric TBE exhibits homeostatic remodeling/repair processes. In these regions, BCs are K5(+)/K14(+), and a subset expresses tissue factor (TF). Passage 1 TBE cells are BCs that are K5(+)/TF+, and half coexpress CD151. Optimal clone formation conditions use an irradiated NIH3T3 fibroblast feeder layer (American Type Culture Collection, Frederick, MD) and serum-supplemented Epicult-Bmedium (Stem cell Technologies, La Jolla, CA). The TF+/CD151(-) BC subpopulation is the most clonogenic BC subtype, and is enriched with K14(+) cells. TF+/CD151(-) BCs generate clones containing BCs that are K5(+)/Trp63(+), but K14(-)/CD151(-). TF+ cells are limited to the clone edge. In conclusion, clonogenic human TBE BCs (1) exhibit a molecular phenotype that is a composite of the normal and remodeling/reparative BC phenotypes observed in tissue, and (2) generate organoid clones that contain phenotypically distinct BC subpopulations.