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At 8 DIV, growth medium was replaced with defined serum-free culture medium consisting of a mixture of a buffered saline solution (10 mM HEPES, pH 7.4, 114 mM NaCl, 26.1 mM NaHCO 3 , 5.3 mM KCl, 1 mM MgCl 2 , 2 mM CaCl 2 , 5 mM glucose, 1 mM glycine, 0.5 mM C 3 H 3 NaO 3 , and 0.001% phenol red) and phosphate-free Eagle's minimum essential medium (9:1 v/v) supplemented with insulin (7.5 g/mL), transferrin (7.5 g/mL), and sodium selenite (7.5 ng/mL) (ITS supplement, Merck Millipore, #I3146)

Analytical Benchmarking: Procedures utilizing validated reference standards, mass spectrometry, and chromatographic verification for laboratory quality control assessments

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For this application, cyclonic filtration is more effective and easier to maintain than sand filtration