Breast Cancer Research and Treatment

, Volume 144, Issue 3, pp 503–517

Primary breast tumor-derived cellular models: characterization of tumorigenic, metastatic, and cancer-associated fibroblasts in dissociated tumor (DT) cultures

  • Katherine Drews-Elger
  • Joeli A. Brinkman
  • Philip Miller
  • Sanket H. Shah
  • J. Chuck Harrell
  • Thiago G. da Silva
  • Zheng Ao
  • Amy Schlater
  • Diana J. Azzam
  • Kathleen Diehl
  • Dafydd Thomas
  • Joyce M. Slingerland
  • Charles M. Perou
  • Marc E. Lippman
  • Dorraya El-Ashry
Preclinical Study

DOI: 10.1007/s10549-014-2887-9

Cite this article as:
Drews-Elger, K., Brinkman, J.A., Miller, P. et al. Breast Cancer Res Treat (2014) 144: 503. doi:10.1007/s10549-014-2887-9

Abstract

Our goal was to establish primary cultures from dissociation of breast tumors in order to provide cellular models that may better recapitulate breast cancer pathogenesis and the metastatic process. Here, we report the characterization of six cellular models derived from the dissociation of primary breast tumor specimens, referred to as “dissociated tumor (DT) cells.” In vitro, DT cells were characterized by proliferation assays, colony formation assays, protein, and gene expression profiling, including PAM50 predictor analysis. In vivo, tumorigenic and metastatic potential of DT cultures was assessed in NOD/SCID and NSG mice. These cellular models differ from recently developed patient-derived xenograft models in that they can be used for both in vitro and in vivo studies. PAM50 predictor analysis showed DT cultures similar to their paired primary tumor and as belonging to the basal and Her2-enriched subtypes. In vivo, three DT cultures are tumorigenic in NOD/SCID and NSG mice, and one of these is metastatic to lymph nodes and lung after orthotopic inoculation into the mammary fat pad, without excision of the primary tumor. Three DT cultures comprised of cancer-associated fibroblasts (CAFs) were isolated from luminal A, Her2-enriched, and basal primary tumors. Among the DT cells are those that are tumorigenic and metastatic in immunosuppressed mice, offering novel cellular models of ER-negative breast cancer subtypes. A group of CAFs provide tumor subtype-specific components of the tumor microenvironment (TME). Altogether, these DT cultures provide closer-to-primary cellular models for the study of breast cancer pathogenesis, metastasis, and TME.

Keywords

Primary cultures Tumors ER-negative breast cancer Metastatic xenograft models 

Abbreviations

DT

Dissociated tumor cells

CAF

Cancer-associated fibroblasts

ERα

Estrogen receptor alpha

TME

Tumor microenvironment

EMT

Epithelial mesenchymal transition

IHC

Immunohistochemistry

FAP

Fibroblast activation protein

α-SMA

Alpha smooth muscle actin

TN

Triple-negative

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Katherine Drews-Elger
    • 1
  • Joeli A. Brinkman
    • 1
  • Philip Miller
    • 1
    • 2
  • Sanket H. Shah
    • 1
    • 2
  • J. Chuck Harrell
    • 3
  • Thiago G. da Silva
    • 1
    • 4
  • Zheng Ao
    • 1
  • Amy Schlater
    • 5
  • Diana J. Azzam
    • 1
  • Kathleen Diehl
    • 6
  • Dafydd Thomas
    • 7
  • Joyce M. Slingerland
    • 1
  • Charles M. Perou
    • 3
  • Marc E. Lippman
    • 1
  • Dorraya El-Ashry
    • 1
  1. 1.Sylvester Comprehensive Cancer Center, Department of MedicineUniversity of Miami Miller School of MedicineMiamiUSA
  2. 2.Sheila and David Fuente Graduate Program in Cancer BiologyUniversity of Miami Miller School of MedicineMiamiUSA
  3. 3.Department of GeneticsUniversity of North Carolina at Chapel HillChapel HillUSA
  4. 4.Department of SurgeryUniversity of Miami Miller School of MedicineMiamiUSA
  5. 5.Division of Hematology/Oncology, Department of Internal MedicineUniversity of Michigan Medical CenterAnn ArborUSA
  6. 6.Department of SurgeryUniversity of Michigan Medical CenterAnn ArborUSA
  7. 7.Department of PathologyUniversity of Michigan Medical CenterAnn ArborUSA

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