Identification of multipotent mesenchymal stromal cells in the reactive stroma of a prostate cancer xenograft by side population analysis

Albert Santamaria-Martínez, Jordi Barquinero, Anna Barbosa-Desongles, Antoni Hurtado, Tomàs Pinós, Joan Seoane, Marie France Poupon, Joan Morote, Jaume Reventós, Francina Munell

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26 Citations (Scopus)


Cancer stem cells are a distinct cellular population that is believed to be responsible for tumor initiation and maintenance. Recent data suggest that solid tumors also contain another type of stem cells, the mesenchymal stem cells or multipotent mesenchymal stromal cells (MSCs), which contribute to the formation of tumor-associated stroma. The Hoechst 33342 efflux assay has proved useful to identify a rare cellular fraction, named Side Population (SP), enriched in cells with stem-like properties. Using this assay, we identified SP cells in a prostate cancer xenograft containing human prostate cancer cells and mouse stromal cells. The SP isolation, subculture and sequential sorting allowed the generation of single-cell-derived clones of murine origin that were recognized as MSC by their morphology, plastic adherence, proliferative potential, adipogenic and osteogenic differentiation ability and immunophenotype (CD45 - , CD81 + and Sca-1 + ). We also demonstrated that SP clonal cells secrete transforming growth factor β1 (TGF-β1) and that their inhibition reduces proliferation and accelerates differentiation. These results reveal the existence of SP cells in the stroma of a cancer xenograft, and provide evidence supporting their MSC nature and the role of TGF-β1 in maintaining their proliferation and undifferentiated status. Our data also reveal the usefulness of the SP assay to identify and isolate MSC cells from carcinomas. © 2009 Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)3004-3013
JournalExperimental Cell Research
Issue number17
Publication statusPublished - 15 Oct 2009


  • Multipotent mesenchymal stromal cell
  • Prostate cancer
  • Side population
  • Stroma
  • Transforming growth factor-β


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