Chapter 26
Inducing Sequential Cycles of Epithelial-Mesenchymal
and Mesenchymal-Epithelial Transitions in Mammary
Epithelial Cells
Cecile Davaine, Eva Hadadi, William Taylor, Annelise Bennaceur-Griscelli,
and Herve ´ Acloque
Abstract
Epithelial-Mesenchymal Transition (EMT) and its reciprocal Mesenchymal-Epithelial Transition (MET)
occur naturally as a cycling process during embryonic and foetal development. The capacity of such iterative
cycles to drive cell fate and cellular and molecular behaviour in physiology or pathology remains unclear. We
describe here a protocol to induce successive cycles of EMT/MET in an untransformed human mammary
epithelial cell line (MCF10A) as well as the necessary controls for cycle validation.
Key words EMT, MET, MCF10A, Epithelial-mesenchymal transition, Mesenchymal-epithelial transition, Cycles
1 Introduction
Epithelial-to-mesenchymal transition (EMT) is a process through
which cells shift their properties, losing strong cell–cell adherence
and gaining migratory capacities [1]. EMT was first described at the
onset of gastrulation in the chick embryo, when cells from the
epiblast delaminate at the primitive streak to form the mesendoderm [2]. Many other developmental processes in amniotes go
through this process, like the prototypic neural crest cell delamination or myotome formation [3–6]. Interestingly, EMT is a reversible process and mesenchymal cells can revert to an epithelial
phenotype to produce new epithelial layers. This phenomenon,
called mesenchymal-epithelial transition (MET), has been observed
during embryogenesis and organogenesis [7]. These round trips
between two antagonistic cellular states allow co-opting the properties of mesenchymal cells (motility, directional migration, individual cell migration) or epithelial cells (polarization, mechanical
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_26,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
341
Inducing Sequential Cycles of Epithelial-Mesenchymal
and Mesenchymal-Epithelial Transitions in Mammary
Epithelial Cells
Cecile Davaine, Eva Hadadi, William Taylor, Annelise Bennaceur-Griscelli,
and Herve ´ Acloque
Abstract
Epithelial-Mesenchymal Transition (EMT) and its reciprocal Mesenchymal-Epithelial Transition (MET)
occur naturally as a cycling process during embryonic and foetal development. The capacity of such iterative
cycles to drive cell fate and cellular and molecular behaviour in physiology or pathology remains unclear. We
describe here a protocol to induce successive cycles of EMT/MET in an untransformed human mammary
epithelial cell line (MCF10A) as well as the necessary controls for cycle validation.
Key words EMT, MET, MCF10A, Epithelial-mesenchymal transition, Mesenchymal-epithelial transition, Cycles
1 Introduction
Epithelial-to-mesenchymal transition (EMT) is a process through
which cells shift their properties, losing strong cell–cell adherence
and gaining migratory capacities [1]. EMT was first described at the
onset of gastrulation in the chick embryo, when cells from the
epiblast delaminate at the primitive streak to form the mesendoderm [2]. Many other developmental processes in amniotes go
through this process, like the prototypic neural crest cell delamination or myotome formation [3–6]. Interestingly, EMT is a reversible process and mesenchymal cells can revert to an epithelial
phenotype to produce new epithelial layers. This phenomenon,
called mesenchymal-epithelial transition (MET), has been observed
during embryogenesis and organogenesis [7]. These round trips
between two antagonistic cellular states allow co-opting the properties of mesenchymal cells (motility, directional migration, individual cell migration) or epithelial cells (polarization, mechanical
Kyra Campbell and Eric Theveneau (eds.), The Epithelial-to Mesenchymal Transition: Methods and Protocols,
Methods in Molecular Biology, vol. 2179, https://doi.org/10.1007/978-1-0716-0779-4_26,
© Springer Science+Business Media, LLC, part of Springer Nature 2021
341