The components of Plasmax are reported in table S1. Stock solutions were prepared in individual tubes for single use and stored at −78°C. To increase the solubility of its components, the pH of solution 1 was adjusted to 1.00 with HCl, and the pH of solution 4 (urate) was adjusted to 13.30 with NaOH. To prepare a bottle of Plasmax, stock solutions 1 to 5 were added to 500 ml of commercially available EBSS (cat. no. 24010043; Thermo Fisher Scientific, MA, USA), in combination with l-glutamine (cat. no. 25030081; Thermo Fisher Scientific), sodium pyruvate (S8636; Sigma-Aldrich, MO, USA), and 2.5% FBS (Thermo Fisher Scientific). After preparation and equilibration with 5% CO2, the pH of different batches of Plasmax was 7.1 to 7.3, with thus not needing for further adjustment. The buffering capacity of Plasmax ensured that its pH remained stable when MDA-MB-468 cells (seeded at 200,000 cells per well) were cultured in 8 ml of medium for 72 hours. The osmolarity of Plasmax was 280 mOsm/kg as measured with a vapor pressure osmometer (Wescor 5500). Trace elements (including sodium selenite; table S1) were supplemented at the concentrations found in Advanced DMEM-F12 (cat. no. 12634028; Thermo Fisher Scientific).

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