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Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque

1
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
2
California National Primate Research Center, University of California, Davis, School of Veterinary Medicine, Davis, CA 95616, USA
*
Author to whom correspondence should be addressed.
Methods Protoc. 2019, 2(4), 79; https://doi.org/10.3390/mps2040079
Received: 26 August 2019 / Revised: 26 September 2019 / Accepted: 27 September 2019 / Published: 1 October 2019
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–liquid 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. View Full-Text
Keywords: rhesus macaque; tracheal basal cells; primary culture; air–liquid interface; tracheospheres rhesus macaque; tracheal basal cells; primary culture; air–liquid interface; tracheospheres
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MDPI and ACS Style

Engler, A.E.; Mostoslavsky, G.; Miller, L.; Rock, J.R. Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque. Methods Protoc. 2019, 2, 79. https://doi.org/10.3390/mps2040079

AMA Style

Engler AE, Mostoslavsky G, Miller L, Rock JR. Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque. Methods and Protocols. 2019; 2(4):79. https://doi.org/10.3390/mps2040079

Chicago/Turabian Style

Engler, Anna E., Gustavo Mostoslavsky, Lisa Miller, and Jason R. Rock 2019. "Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque" Methods and Protocols 2, no. 4: 79. https://doi.org/10.3390/mps2040079

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