Abstract
Microenvironmental mechanical signals are fundamental regulators of cell behavior both in physiological and in pathological context, particularly in the induction and maintenance of tumorigenic properties. It is thus of utmost importance to experimentally recreate conditions that mimic the physical attributes of real tissues to study their impact on cell behavior and in particular on the induction of cancer stem cell (CSC) properties. Here we present protocols to investigate the role of mechanical stiffness on reprogramming of primary mammary gland cells into CSCs, including the synthesis of hydrogel substrates of the desired stiffness, the isolation and culture of primary differentiated normal cells derived from the human mammary gland, and the assessment of their CSC attributes after oncogene-mediated transformation.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Eyckmans J, Boudou T, Yu X, Chen CS (2011) A hitchhiker’s guide to mechanobiology. Dev Cell 21(1):35–47
Humphrey JD, Dufresne ER, Schwartz MA (2014) Mechanotransduction and extracellular matrix homeostasis. Nat Rev Mol Cell Biol 15(12):802–812
Panciera T, Azzolin L, Cordenonsi M, Piccolo S (2017) Mechanobiology of YAP and TAZ in physiology and disease. Nat Rev Mol Cell Biol 18(12):758–770
Discher DE, Mooney DJ, Zandstra PW (2009) Growth factors, matrices, and forces combine and control stem cells. Science 324(5935):1673–1677
Vining KH, Mooney DJ (2017) Mechanical forces direct stem cell behaviour in development and regeneration. Nat Rev Mol Cell Biol 18(12):728–742
Northey JJ, Przybyla L, Weaver VM (2017) Tissue force programs cell fate and tumor aggression. Cancer Discov 7(11):1224–1237
Piccolo S, Panciera T, Contessotto P, Cordenonsi M (2023) YAP/TAZ as master regulators in cancer: modulation, function and therapeutic approaches. Nat Cancer 4(1):9–26
Visvader JE, Lindeman GJ (2008) Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer 8(10):755–768
Gupta PB, Chaffer CL, Weinberg RA (2009) Cancer stem cells: mirage or reality? Nat Med 15(9):1010–1012
Brusatin G, Panciera T, Gandin A, Citron A, Piccolo S (2018) Biomaterials and engineered microenvironments to control YAP/TAZ-dependent cell behaviour. Nat Mater 17(12):1063–1075
Schwartz MA, Chen CS (2013) Cell biology. Deconstructing dimensionality. Science 339(6118):402–404
Panciera T, Citron A, Di Biagio D, Battilana G, Gandin A, Giulitti S et al (2020) Reprogramming normal cells into tumour precursors requires ECM stiffness and oncogene-mediated changes of cell mechanical properties. Nat Mater 19(7):797–806
Van Keymeulen A, Lee MY, Ousset M, Brohee S, Rorive S, Giraddi RR et al (2015) Reactivation of multipotency by oncogenic PIK3CA induces breast tumour heterogeneity. Nature 525(7567):119–123
Linnemann JR, Miura H, Meixner LK, Irmler M, Kloos UJ, Hirschi B et al (2015) Quantification of regenerative potential in primary human mammary epithelial cells. Development 142(18):3239–3251
Acerbi I, Cassereau L, Dean I, Shi Q, Au A, Park C et al (2015) Human breast cancer invasion and aggression correlates with ECM stiffening and immune cell infiltration. Integr Biol (Camb) 7(10):1120–1134
Gandin A, Torresan V, Ulliana L, Panciera T, Contessotto P, Citron A et al (2022) Broadly applicable hydrogel fabrication procedures guided by YAP/TAZ-activity reveal stiffness, adhesiveness, and nuclear projected area as checkpoints for Mechanosensing. Adv Healthc Mater 11(3):e2102276
Damljanovic V, Lagerholm BC, Jacobson K (2005) Bulk and micropatterned conjugation of extracellular matrix proteins to characterized polyacrylamide substrates for cell mechanotransduction assays. BioTechniques 39(6):847–851
Panciera T, Azzolin L, Fujimura A, Di Biagio D, Frasson C, Bresolin S et al (2016) Induction of expandable tissue-specific stem/progenitor cells through transient expression of YAP/TAZ. Cell Stem Cell 19(6):725–737
Acknowledgements
This work was supported by a PRIN 2017 grant (no. 2017L8FWY8_004) and a CARIPARO Starting Grant (no. C94I19001680001) to T.P.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Gandin, A., Contessotto, P., Panciera, T. (2024). Methods to Study the Role of Mechanical Signals in the Induction of Cancer Stem Cells. In: Papaccio, F., Papaccio, G. (eds) Cancer Stem Cells. Methods in Molecular Biology, vol 2777. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3730-2_13
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3730-2_13
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3729-6
Online ISBN: 978-1-0716-3730-2
eBook Packages: Springer Protocols