title: Primary breast tumor-derived cellular models: characterization of tumorigenic, metastatic, and cancer-associated fibroblasts in dissociated tumor (DT) cultures.
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.
authors.affiliation: Sylvester Comprehensive Cancer Center, Department of Medicine, University of Miami Miller School of Medicine, Biomedical Research Building, 1501 NW 10th Avenue, Miami, FL, 33136, USA.
authors.forename: Katherine
authors.initials: K
authors.lastname: Drews-Elger
authors.forename: Joeli A
authors.initials: JA
authors.lastname: Brinkman
authors.forename: Philip
authors.initials: P
authors.lastname: Miller
authors.forename: Sanket H
authors.initials: SH
authors.lastname: Shah
authors.forename: J Chuck
authors.initials: JC
authors.lastname: Harrell
authors.forename: Thiago G
authors.initials: TG
authors.lastname: da Silva
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ao
authors.forename: Amy
authors.initials: A
authors.lastname: Schlater
authors.forename: Diana J
authors.initials: DJ
authors.lastname: Azzam
authors.forename: Kathleen
authors.initials: K
authors.lastname: Diehl
authors.forename: Dafydd
authors.initials: D
authors.lastname: Thomas
authors.forename: Joyce M
authors.initials: JM
authors.lastname: Slingerland
authors.forename: Charles M
authors.initials: CM
authors.lastname: Perou
authors.forename: Marc E
authors.initials: ME
authors.lastname: Lippman
authors.forename: Dorraya
authors.initials: D
authors.lastname: El-Ashry
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002453:Cell Cycle; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic; D004195:Disease Models, Animal; D005260:Female; D005347:Fibroblasts / Q000473:pathology; D020869:Gene Expression Profiling; D064593:Heterografts; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D009362:Neoplasm Metastasis; D061251:Primary Cell Culture / Q000379:methods; D047368:Tumor Burden; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
chemical_list: D014408:Biomarkers, Tumor