Cancer Stem Cell Migration in Three-Dimensional Aligned Collagen Matrices

Arja Ray, Rachel K. Morford, Paolo P. Provenzano

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Cell migration is strongly influenced by the organization of the surrounding 3-D extracellular matrix. In particular, within fibrous solid tumors, carcinoma cell invasion may be directed by patterns of aligned collagen in the extra-epithelial space. Thus, studying the interactions of heterogeneous populations of cancer cells that include the stem/progenitor-like cancer stem cell subpopulation and aligned collagen networks is critical to our understanding of carcinoma dissemination. Here, we describe a robust method to generate aligned collagen matrices in vitro that mimic in vivo fiber organization. Subsequently, a protocol is presented for seeding aligned matrices with distinct carcinoma cell subpopulations and performing live cell imaging and quantitative analysis of cell migration. Together, the engineered constructs and the imaging techniques laid out here provide a platform to study cancer stem cell migration in 3-D anisotropic collagen with real-time visualization of cellular interactions with the fibrous matrix.

Original languageEnglish (US)
Article numbere57
JournalCurrent Protocols in Stem Cell Biology
Issue number1
StatePublished - Aug 2018

Bibliographical note

Funding Information:
P.P.P. and this work were supported by Research Scholar Grant RSG-14-171-01-CSM from the American Cancer Society. This work was also supported by the NIH (R01CA181385 to P.P.P. and U54CA210190 University of Minnesota Physical Sciences in Oncology Center Project 2 to P.P.P.), UMN College of Science and Engineering (P.P.P.), and Masonic Cancer Center (P.P.P.), and grants from the UMN Institute for Engineering in Medicine (P.P.P.) and the Randy Shaver Research and Community Fund (P.P.P.). A.R. was supported by a UMN Doctoral Dissertation Fellowship. R.K.M. is supported by NSF graduate research fellowship 00039202. The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH or other funding agencies. We thank members of the Proven-zano laboratory for insightful comments regarding this work, the Tranquillo lab for lending equipment and expertise, and the Odde lab for sharing knowledge on time-lapse imaging and cell migration analysis.

Publisher Copyright:
© 2018 John Wiley & Sons, Inc.


  • 3-D collagen matrices
  • cancer stem cell
  • cell migration
  • collagen alignment


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