Improving the metabolic fidelity of cancer models with a physiological cell culture medium

Sci Adv. 2019 Jan 2;5(1):eaau7314. doi: 10.1126/sciadv.aau7314. eCollection 2019 Jan.


Currently available cell culture media may not reproduce the in vivo metabolic environment of tumors. To demonstrate this, we compared the effects of a new physiological medium, Plasmax, with commercial media. We prove that the disproportionate nutrient composition of commercial media imposes metabolic artifacts on cancer cells. Their supraphysiological concentrations of pyruvate stabilize hypoxia-inducible factor 1α in normoxia, thereby inducing a pseudohypoxic transcriptional program. In addition, their arginine concentrations reverse the urea cycle reaction catalyzed by argininosuccinate lyase, an effect not observed in vivo, and prevented by Plasmax in vitro. The capacity of cancer cells to form colonies in commercial media was impaired by lipid peroxidation and ferroptosis and was rescued by selenium present in Plasmax. Last, an untargeted metabolic comparison revealed that breast cancer spheroids grown in Plasmax approximate the metabolic profile of mammary tumors better. In conclusion, a physiological medium improves the metabolic fidelity and biological relevance of in vitro cancer models.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arginine / metabolism
  • Argininosuccinate Lyase / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Culture Media*
  • Female
  • Ferroptosis / drug effects
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Lipid Peroxidation / drug effects
  • Models, Biological*
  • Pyruvic Acid / metabolism
  • Sodium Selenite / pharmacology
  • Spheroids, Cellular / metabolism
  • Triple Negative Breast Neoplasms / metabolism*
  • Triple Negative Breast Neoplasms / pathology*
  • Tumor Microenvironment / physiology*
  • Urea / metabolism


  • Culture Media
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Pyruvic Acid
  • Urea
  • Arginine
  • Argininosuccinate Lyase
  • Sodium Selenite