Assembly of lung progenitors into developmentally-inspired geometry drives differentiation via cellular tension

John P Soleas; Elisa D'Arcangelo; Linwen Huang; Golnaz Karoubi; Maria Cristina Nostro; Alison P McGuigan; Thomas K Waddell

doi:10.1016/j.biomaterials.2020.120128

Assembly of lung progenitors into developmentally-inspired geometry drives differentiation via cellular tension

Biomaterials. 2020 Sep:254:120128. doi: 10.1016/j.biomaterials.2020.120128. Epub 2020 May 20.

Authors

John P Soleas¹, Elisa D'Arcangelo², Linwen Huang², Golnaz Karoubi³, Maria Cristina Nostro⁴, Alison P McGuigan⁵, Thomas K Waddell⁶

Affiliations

¹ Institute for Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada, M5S 3G9; Latner Thoracic Surgery Research Laboratories, Toronto General Hospital, 101 College St., Toronto, ON, M5G 1L7, Canada.
² Institute for Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada, M5S 3G9.
³ Institute for Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada, M5S 3G9; Department of Mechanical and Industrial Engineering, University of Toronto, 5 King's College Rd., Toronto, ON, M5S 3G8, Canada.
⁴ McEwen Stem Cell Institute, University Health Network, 101 College St., Toronto, ON, M5G 1L7, Canada; Department of Physiology, University of Toronto, 1 King's College Circle, Toronto, ON, M5S 1A8, Canada.
⁵ Institute for Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada, M5S 3G9; Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College St., Toronto, ON, M5S 3E5, Canada. Electronic address: alison.mcguigan@utoronto.ca.
⁶ Institute for Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada, M5S 3G9; Latner Thoracic Surgery Research Laboratories, Toronto General Hospital, 101 College St., Toronto, ON, M5G 1L7, Canada; Institute of Medical Science, University of Toronto, 1 King's College Circle, Toronto, ON, M5S 1A8, Canada. Electronic address: tom.waddell@uhn.ca.

PMID: 32474250
DOI: 10.1016/j.biomaterials.2020.120128

Abstract

During organogenesis groups of differentiating cells self-organize into a series of structural intermediates with defined architectural forms. Evidence is emerging that such architectural forms are important in guiding cell fate, yet in vitro methods to guide cell fate have focused primarily on un-patterned exposure of stems cells to developmentally relevant chemical cues. We set out to ask if organizing differentiating lung progenitors into developmentally relevant structures could be used to influence differentiation status. Specifically, we use elastomeric substrates to guide self-assembly of human pluripotent stem cell-derived lung progenitors into developmentally-relevant sized tubes and assess the impact on differentiation. Culture in 100 μm tubes reduced the percentage of SOX2⁺SOX9⁺ cells and reduced proximal fate potential compared to culture in 400 μm tubes or on flat surfaces. Cells in 100 μm tubes curved to conform to the tube surface and experienced increased cellular tension and reduced elongation. Pharmacologic disruption of tension through inhibition of ROCK, myosin II activity and actin polymerization in tubes resulted in maintenance of SOX2⁺SOX9⁺ populations. Furthermore, this effect required canonical WNT signaling. This data suggests that structural forms, when developmentally relevant, can drive fate choice during directed differentiation via a tension-based canonical WNT dependent mechanism.

Keywords: Directed differentiation; Lung; Morphogenesis; Stem cell differentiation; Tissue architecture; Tissue engineering.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Differentiation
Humans
Lung
Pluripotent Stem Cells*
Wnt Signaling Pathway

Grants and funding

Canadian Institute for Health Reseach