Journal
AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY
Volume 49, Issue 6, Pages 1127-1134Publisher
AMER THORACIC SOC
DOI: 10.1165/rcmb.2013-0049OC
Keywords
basal cell; remodeling; clonogenic frequency; phenotypic plasticity; stem cell
Funding
- National Institutes of Health [RC1 HL099461, RO1 HL075585, HL075585-S1, K18-HL107686, P30 CA 046934]
- Young Clinical Scientist Award-Flight Attendants Medical Research Institute [ES014448, KL2RR025779]
- CounterACT Program, the Office of the Director at the National Institutes of Health
- National Institute of Environmental Health Sciences [U54 ES015678]
- Max and Yetta Karasik Family Foundation
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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.
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