Cell tak cell and tissue adhesive

Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) of CD4⁺ T cells were measured using a Seahorse XFe96 Analyzers. For common tests, prepared cultured cells or in vivo isolated cells in XF RPMI medium supplemented with 10 mM glucose, 1 mM pyruvate and 2 mM L-glutamine, added 2 × 10⁵ (in vitro cultured for 24 h) or 3 × 10⁵ (fresh isolated from in vivo) T cells into each well of Seahorse XF96 cell culture microplates (Coated with 22.4 μg/ml Corning® Cell-Tak™ Cell and Tissue Adhesive), spined down and preincubated at 37°C for around 30 min in the absence of CO₂, then ran the program of the Seahorse XF Cell Mito Stress Test. For the measurement of basal glycolysis of T cells cultured with 5.5 mM, 25 mM and 50 mM glucose, cells were harvested after 24 h culture in indicate concentrations of glucose or galactose, then prepared the cells in XF RPMI medium supplemented with 1 mM pyruvate, 2 mM L-glutamine and indicate concentrations of glucose or galactose (Same concentrations with T cell culture), then preincubated and measured the basal ECAR.