Life-long functional regeneration of in vivo airway epithelium by the engraftment of airway basal stem cells.

Autor: Ma L; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA.; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA, USA., Thapa BR; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA.; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA, USA.; Department of Biology, Boston University, Boston, MA, USA., Le Suer JA; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA.; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA, USA., Tilston-Lünel A; Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN, USA., Herriges MJ; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA.; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA, USA., Wang F; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA., Bawa PS; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA., Varelas X; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA.; Department of Biochemistry and Cell Biology, Boston University School of Medicine, Boston, MA, USA., Hawkins FJ; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA.; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA, USA., Kotton DN; Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, MA, USA. dkotton@bu.edu.; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA, USA. dkotton@bu.edu.
Jazyk: angličtina
Zdroj: Nature protocols [Nat Protoc] 2024 Nov 05. Date of Electronic Publication: 2024 Nov 05.
DOI: 10.1038/s41596-024-01067-y
Abstrakt: Durable and functional regeneration of the airway epithelium in vivo with transplanted stem cells has the potential to reconstitute healthy tissue in diseased airways, such as in cystic fibrosis or primary ciliary dyskinesia. Here, we present detailed protocols for the preparation and culture expansion of murine primary and induced pluripotent stem cell-derived airway basal stem cells (iBCs) and methods for their intra-airway transplantation into polidocanol-conditioned murine recipients to achieve durable in vivo airway regeneration. Reconstitution of the airway tissue resident epithelial stem cell compartment of immunocompetent mice with syngeneic donor cells leverages the extensive self-renewal and multipotent differentiation properties of basal stem cells (BCs) to durably generate a broad diversity of mature airway epithelial lineages in vivo. Engrafted donor-derived cells re-establish planar cell polarity as well as functional ciliary transport. By using this same approach, human primary BCs or iBCs transplanted into NOD-SCID gamma recipient mice similarly display engraftment and multilineage airway epithelial differentiation in vivo. The time to generate mouse or human iBCs takes ~60 d, which can be reduced to ~20 d if previously differentiated cells are thawed from cryopreserved iBC archives. The tracheal conditioning regimen and cell transplantation procedure is completed in 1 d. A competent graduate student or postdoctoral trainee should be able to perform the procedures listed in this protocol.
(© 2024. Springer Nature Limited.)
Databáze: MEDLINE
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