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Basal cells are known to self-renew and differentiate into airway ciliated and secretory cells. However, it is not clear if every basal cell functions as a stem cell. To address functional heterogeneity amongst the basal cell population, we developed a novel monoclonal antibody, HLO1-6H5, that identifies a subset of KRT5+ (cytokeratin 5) basal cells. We used HLO1-6H5 and other known basal cell-reactive reagents to isolate viable airway subsets from primary human airway epithelium by Fluorescence Activated Cell Sorting. Isolated primary cell subsets were assessed for the stem cell capabilities of self-renewal and differentiation in the bronchosphere assay, which revealed that bipotent stem cells were, at minimum 3-fold enriched in the HLO1-6H5+ cell subset. Crosslinking-mass spectrometry identified the HLO1-6H5 target as a glycosylated TFRC\/CD71 (transferrin receptor) proteoform. The HLO1-6H5 antibody provides a valuable new tool for identifying and isolating a subset of primary human airway basal cells that are substantially enriched for bipotent stem\/progenitor cells and reveals TFRC as a defining surface marker for this novel cell subset.<\/jats:p>","DOI":"10.3390\/ijms25189863","type":"journal-article","created":{"date-parts":[[2024,9,12]],"date-time":"2024-09-12T08:04:09Z","timestamp":1726128249000},"page":"9863","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Identification of a Novel Subset of Human Airway Epithelial Basal Stem Cells"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7890-8287","authenticated-orcid":false,"given":"Christopher","family":"Cheng","sequence":"first","affiliation":[{"name":"Oregon Stem Cell Center, Pap\u00e8 Family Pediatric Research Institute, Department of Pediatrics, Oregon Health and Science University, Portland, OR 97239-3098, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parul","family":"Katoch","sequence":"additional","affiliation":[{"name":"Cancer Early Detection Advanced Research Center, Knight Cancer Institute, Oregon Health and Science University, Portland, OR 97239-3098, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong-Ping","family":"Zhong","sequence":"additional","affiliation":[{"name":"Oregon Stem Cell Center, Pap\u00e8 Family Pediatric Research Institute, Department of Pediatrics, Oregon Health and Science University, Portland, OR 97239-3098, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire T.","family":"Higgins","sequence":"additional","affiliation":[{"name":"Oregon Stem Cell Center, Pap\u00e8 Family Pediatric Research Institute, Department of Pediatrics, Oregon Health and Science University, Portland, OR 97239-3098, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Moredock","sequence":"additional","affiliation":[{"name":"Oregon Stem Cell Center, Pap\u00e8 Family Pediatric Research Institute, Department of Pediatrics, Oregon Health and Science University, Portland, OR 97239-3098, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5859-388X","authenticated-orcid":false,"given":"Matthew E. 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