Adaptation of breast cancer cells to serum-free defined medium reveals changes in proliferation, adhesion, and lipid metabolism

MDA-MB-231, SK-BR-3, and BT-474 breast cancer cells show distinct adaptation to serum-free defined medium, with cholesterol-homeostasis gene programs enriched and SREBF2/HMGCR/LDLR upregulated across all three lines.

• Why it matters: Serum-free, physiologically relevant culture can reveal metabolic vulnerabilities that are masked by non-physiological nutrient levels and poorly defined serum components.
• What they did: The authors stepwise reduced fetal bovine serum (FBS) to serum-free defined medium (DM) and measured proliferation, adhesion, morphology, and transcriptional changes using RNA sequencing in HER2-positive SK-BR-3, triple-negative MDA-MB-231, and ER-positive/HER2-positive BT-474 cells.
• The result: MDA-MB-231 maintained proliferation and adhesion without exogenous coating, SK-BR-3 sustained proliferation but needed a serum-derived coating for adhesion, and BT-474 failed after complete serum withdrawal but proliferated with 1% FBS; cholesterol homeostasis gene sets were enriched in SK-BR-3 and MDA-MB-231, and cholesterol regulatory genes increased across all three lines.

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