Selection, establishment and characterization of cell lines derived from a chemically-induced rat mammary heterogeneous tumor, by flow cytometry, transmission electron microscopy, and immunohistochemistry

Laura Teodori, Fausto Tagliaferri, Francesco Stipa, Maria Giovanna Valente, Dario Coletti, Angelo Manganelli, Mario Guglielmi, Luciana Santoro D'Angelo, Hartmut Schäfer, Wolfgang Göhde

Research output: Contribution to journalArticlepeer-review

Abstract

In order to isolate, characterize, and establish culture cell lines with different diagnostic and prognostic significance, derived from multiclonal neoplasms, a ductal infiltrating mammary tumor was induced in rats by 7,12- dimethylbenz[a]anthracene. Clones with different DNA/protein content, being the DI of 1.16, 1.30, and 1.60, respectively, were observed in the primary tumor. Biparametric flow cytometry suggested that the clone at 1.30 is made up of two subpopulations with different protein and slightly different DNA contents. The culture, after a few passages, exhibited the presence of aneuploid cells and the absence of diploid components, demonstrating that only tumor cells survived. The limiting dilution method gave rise to four lines with DI of 1.16, 1.25, 1.30, and 1.50; a mean chromosome number of 45, 46, 47, and 88, respectively; and different morphological and ultrastructural features. These characteristics were stable during the experimental procedure, that is, for about 20 passages. Conversely, the detection of cytoskeletal proteins indicated that the tumor epithelial cells underwent early dedifferentiation into sarcoma-like cells showing markers of stromal cell type and thus exhibiting phenotypic instability in vitro, a feature reported in many advanced human breast cancers in vivo. In conclusion, this cellular model represents the in vivo situation and appears suitable for in vitro studies of tumor cell characteristics and might be used to predict clinical behavior.

Original languageEnglish
Pages (from-to)153-162
Number of pages10
JournalIn Vitro Cellular & Developmental Biology - Animal
Volume36
Issue number3
DOIs
Publication statusPublished - Mar 2000

Keywords

  • Breast cancer
  • DNA content
  • Flow cytometry
  • Rat cell lines
  • Tumor heterogeneity

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

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