Format

Send to

Choose Destination
Cell Stem Cell. 2016 Aug 4;19(2):217-231. doi: 10.1016/j.stem.2016.05.012. Epub 2016 Jun 16.

Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells.

Author information

1
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Division of Pediatric Pulmonary Medicine, Massachusetts General Hospital for Children, Boston, MA 02114, USA.
2
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Massachusetts General Hospital, Boston, MA 02114, USA.
3
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
4
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Division of Pediatric Pulmonary Medicine, Massachusetts General Hospital for Children, Boston, MA 02114, USA.
5
Department of Anatomy and Cell Biology, College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
6
Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Stem Cell Institute, 7 Divinity Avenue, Cambridge, MA 02138, USA.
7
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35233, USA; Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35233, USA.
8
Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL 35233, USA.
9
CFFT Lab, Cystic Fibrosis Foundation Therapeutics, Lexington, MA 01730, USA.
10
Division of Pediatric Pulmonary Medicine, Massachusetts General Hospital for Children, Boston, MA 02114, USA.
11
Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Massachusetts General Hospital, Boston, MA 02114, USA.
12
Departments of Medicine and Biochemistry, Stanford University, Stanford, CA 94305, USA.
13
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35233, USA; Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL 35233, USA; Department of Cell, Developmental, and Integrative Biology, University of Alabama at Birmingham, AL 35294, USA; Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35233, USA.
14
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Division of Pediatric Pulmonary Medicine, Massachusetts General Hospital for Children, Boston, MA 02114, USA; Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Division of Otology and Laryngology, Massachusetts Eye and Ear, Boston, MA 02114, USA. Electronic address: jrajagopal@partners.org.

Abstract

Functional modeling of many adult epithelia is limited by the difficulty in maintaining relevant stem cell populations in culture. Here, we show that dual inhibition of SMAD signaling pathways enables robust expansion of primary epithelial basal cell populations. We find that TGFβ/BMP/SMAD pathway signaling is strongly activated in luminal and suprabasal cells of several epithelia, but suppressed in p63+ basal cells. In airway epithelium, SMAD signaling promotes differentiation, and its inhibition leads to stem cell hyperplasia. Using dual SMAD signaling inhibition in a feeder-free culture system, we have been able to expand airway basal stem cells from multiple species. Expanded cells can produce functional airway epithelium physiologically responsive to clinically relevant drugs, such as CFTR modulators. This approach is effective for the clonal expansion of single human cells and for basal cell populations from epithelial tissues from all three germ layers and therefore may be broadly applicable for modeling of epithelia.

KEYWORDS:

TGFβ/BMP4/SMAD signaling; dedifferentiation; differentiation; dual SMAD signaling inhibition; epithelial basal and stems cells; p63(+) basal cells; replicative exhaustion; senescence; stemness; telomeres

PMID:
27320041
PMCID:
PMC4975684
DOI:
10.1016/j.stem.2016.05.012
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center