Collagen gel contraction serves to rapidly distinguish epithelial- and mesenchymal-derived cells irrespective of α-smooth muscle actin expression

Helga Lind Nielsen, Thorarinn Gudjonsson, René Villadsen, Lone Rønnov-Jessen, Ole William Petersen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Mesenchymal-like cells in the stroma of breast cancer may arise as a consequence of plasticity within the epithelial compartment, also referred to as epithelial-mesenchymal transition, or by recruitment of genuine mesenchymal cells from the peritumoral stroma. Cells of both the epithelial compartment and the stromal compartment express α smooth muscle actin (α-sm actin) as part of a myoepithelial or a myofibroblastic differentiation program, respectively. Moreover, because both epithelial- and mesenchymal-derived cells are nontumorigenic, other means of discrimination are warranted. Here, we describe the contraction of hydrated collagen gels as a rapid functional assay for the distinction between epithelial- and mesenchymal-derived stromal-like cells irrespective of the status of α-sm actin expression. Three epithelial-derived cell lines and three genuine mesenchymal-derived breast cell lines were plated on top of hydrated collagen lattices. Reduction in gel height was measured every hour for 6 h and after 22 h using an x-y-z automated position table. Significantly, the epithelial-derived cells, irrespective of a high α-sm actin expression, had a fivefold lower contractility (10,0% reduction in gel height) than their true mesenchymal counterparts (53.1% reduction in gel height). To test whether at all force generation could be induced in the nonmesenchymal cells by α-sm actin, transductions were performed to obtain a tetracycline-dependent expression. Expression under these conditions did not augment contractility. It is concluded that epithelial-derived mesenchymal-like cells are functionally defective within a connective tissue environment irrespective of an apparent contractile phenotype.

Original languageEnglish
Pages (from-to)297-303
Number of pages7
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume39
Issue number7
DOIs
Publication statusPublished - Jul 2003

Bibliographical note

Funding Information:
We thank Tove Marianne Lund and Winnie Hansen for expert technical assistance and Keld Ottosen for printing the micrographs. We are gratetul to The Aasted Clinic, the Private Clinic, and the Sr Plastic Surgel~r Clinic for providing the biopsy material. This research was sponsored by Dansk Krzeffforskningsfond, The Danish Research Council, The Novo Nordisk Foundation, The Thaysen Foundation, Friis Fonden, The Meyer Foundation, The Danish Cancer Society, Weimanns Legat (to L. R.-J.), and The U.S. National Cancer Institute (grant CA-64786-02 to Mina J. Bissell and O. W. E).

Other keywords

  • α-sm actin expression
  • Breast
  • Contractility
  • Epithelial-derived mesenchymal cells

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