T84 Cells

Culture medium: Ham's F12

w: 1.0 mM stable Glutamine

w: 1.0 mM Sodium pyruvate

w: 1.1 g/L NaHCO3 (Cytion article number 820600a). Supplements: Supplement the medium with 10% FBS. Dissociation reagent: Accutase. Seeding density: 1 to 2 × 10⁴ cells/cm² (maintain at minimum 1/4 confluency to preserve tight junction phenotype). Freeze medium: As a cryopreservation medium

we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability

or CM-1 (Cytion catalog number 800100)

which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. Incubation atmosphere: 37°C

5% CO2

humidified atmosphere.