{
  "abstract": "Cancer cell metabolic dependencies are strongly influenced by nutrient availability, yet conventional culture conditions contain non-physiological nutrient concentrations and poorly defined serum components that may obscure metabolic vulnerabilities. Here, we investigated the adaptation of three breast cancer cell lines to physiologically relevant, serum-free defined medium (DM). Using a stepwise reduction in fetal bovine serum (FBS), we evaluated proliferation, adhesion, morphology, and transcriptional changes in HER2-positive SK-BR-3, triple-negative MDA-MB-231, and ER-positive/HER2-positive BT-474 cells. The three cell lines differed substantially in their ability to adapt to serum-free conditions. MDA-MB-231 cells adapted readily, maintaining proliferation and adhesion without exogenous substrate coatings. SK-BR-3 cells also achieved sustained proliferation in serum-free DM, but required a serum-derived coating to maintain adhesion. In contrast, BT-474 cells failed to sustain adhesion and proliferation following complete serum withdrawal, although they remained proliferative in medium containing 1% FBS. Adapted cells retained characteristic HER2 or vimentin expression despite changes in morphology and proliferation. RNA sequencing revealed extensive transcriptional remodeling, including reduced expression of cell-cycle-associated programs in SK-BR-3 cells and enrichment of cholesterol homeostasis gene sets in SK-BR-3 and MDA-MB-231 cells. Expression of cholesterol regulatory genes, including SREBF2, HMGCR, and LDLR, increased across all three lines. Together, these findings highlight cell-line-specific adhesion requirements and shared metabolic responses to serum withdrawal, establishing a framework for developing defined culture systems to investigate cancer metabolic dependencies through genetic screening.",
  "abstract_shown": true,
  "affiliations": [
    "Fred Hutchinson Cancer Center"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 3,
  "authors": [
    {
      "name": "C. Aarrestad"
    },
    {
      "name": "T. O. Khatib"
    },
    {
      "affs": [
        0
      ],
      "name": "J. V. Alvarez"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/7gqqjz7e0trnp.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.11.02.762606",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.11.02.762606v1.full",
  "id": "7gqqjz7e0trnp",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.11.02.762606v1",
  "language": null,
  "licence": "CC-BY-ND",
  "licence_label": "CC BY-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "cancer biology"
    ]
  ],
  "openalex": null,
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.11.02.762606v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-11-02",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/7gqqjz7e0trnp.ris",
  "sources": [
    {
      "first_seen": "2026-11-04T12:21:05Z",
      "native_id": "10.64898/2026.11.02.762606",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.11.02.762606v1"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "biorxiv:cancer biology",
      "subfield": "oncology"
    }
  ],
  "title": "Adaptation of breast cancer cells to serum-free defined medium reveals changes in proliferation, adhesion, and lipid metabolism",
  "title_text": "Adaptation of breast cancer cells to serum-free defined medium reveals changes in proliferation, adhesion, and lipid metabolism",
  "type": "preprint",
  "updated_at": "2026-11-06T00:20:51Z",
  "url": "https://research.draconitic.com/paper/7gqqjz7e0trnp",
  "venue": "bioRxiv",
  "versions": []
}