{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:47:48Z","timestamp":1760240868421,"version":"build-2065373602"},"reference-count":10,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,10,1]],"date-time":"2019-10-01T00:00:00Z","timestamp":1569888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000009","name":"Foundation for the National Institutes of Health","doi-asserted-by":"publisher","award":["R01 HL127002","U01 HL134766"],"award-info":[{"award-number":["R01 HL127002","U01 HL134766"]}],"id":[{"id":"10.13039\/100000009","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["MPs"],"abstract":"<jats:p>In this report, we describe methodologies for the isolation and culture of primary rhesus macaque tracheal basal cells, their cryopreservation, long term storage and differentiation. These are comparable to state-of-the-art protocols that have been developed for mouse and human airway basal cells. This method is based on the use of proprietary media, providing an easily reproducible and applicable protocol for usage in biosafety level 2 (BSL2) settings. Tracheas from rhesus macaques were isolated after animal euthanasia and subjected to enzymatic digestion overnight. Cells of the epithelial layer were scraped off of the trachea for cell culture. Twenty-four hours after plating basal cells had attached and nonadherent cells were removed. First passages of basal cells can be frozen for early passage storage in liquid nitrogen or propagated and differentiated on an air\u2013liquid interface and in a tracheosphere assay up to passage seven. This protocol provides a platform for the analysis of basal cells from a close evolutionary relative to humans.<\/jats:p>","DOI":"10.3390\/mps2040079","type":"journal-article","created":{"date-parts":[[2019,10,1]],"date-time":"2019-10-01T11:11:16Z","timestamp":1569928276000},"page":"79","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque"],"prefix":"10.3390","volume":"2","author":[{"given":"Anna E.","family":"Engler","sequence":"first","affiliation":[{"name":"Center for Regenerative Medicine of Boston University and Boston Medical Center, The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gustavo","family":"Mostoslavsky","sequence":"additional","affiliation":[{"name":"Center for Regenerative Medicine of Boston University and Boston Medical Center, The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7557-1316","authenticated-orcid":false,"given":"Lisa","family":"Miller","sequence":"additional","affiliation":[{"name":"California National Primate Research Center, University of California, Davis, School of Veterinary Medicine, Davis, CA 95616, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason R.","family":"Rock","sequence":"additional","affiliation":[{"name":"Center for Regenerative Medicine of Boston University and Boston Medical Center, The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,10,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1146\/annurev-cellbio-100109-104040","article-title":"Epithelial Progenitor Cells in Lung Development, Maintenance, Repair, and Disease","volume":"27","author":"Rock","year":"2011","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"12771","DOI":"10.1073\/pnas.0906850106","article-title":"Basal cells as stem cells of the mouse trachea and human airway epithelium","volume":"106","author":"Rock","year":"2009","journal-title":"Proc. Natl. Acad. Sci. 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Cell"},{"key":"ref_6","unstructured":"Fulcher, M.L., Gabriel, S., Burns, K.A., Yankaskas, J.R., and Randell, S.H. (2005). Well-differentiated human airway epithelial cell cultures. Human Cell Culture Protocols, Humana Press."},{"key":"ref_7","unstructured":"Robert, E.D., and Kuramoto, K. (2005). Sources and methods to obtain stem and progenitor cells Large Animal Models for Stem and Progenitor Cell Analysis. Blood, 22A."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1159\/000459794","article-title":"Transmission of Hepatitis B to the Rhesus Monkey","volume":"7","author":"Scalise","year":"1978","journal-title":"J. Med. Primatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1513\/pats.200805-041HR","article-title":"Airway Epithelial Cells: Current Concepts and Challenges","volume":"5","author":"Crystal","year":"2008","journal-title":"Proc. Am. Thorac. Soc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"16406","DOI":"10.1073\/pnas.0608130103","article-title":"Complete correction of murine Artemis immunodeficiency by lentiviral vector-mediated gene transfer","volume":"103","author":"Mostoslavsky","year":"2006","journal-title":"Proc. Natl. Acad. Sci. 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