P450 glo cyp3a4 assay with luciferin ipa

Samples were taken from the platform modules 17, 24 and 48 hours after each medium change. CC10 production from airway tissue was measured by ELISA (R&D Systems, Minneapolis, MN) performed according to the manufacturer’s specifications. Comparisons to static airway models were made for on-platform airway models at the t=17hr time point only, pre-interaction. Total albumin from liver co-cultures was measured using a human albumin ELISA kit (Bethyl Labs, Montgomery, TX). At the termination of the experiment, CYP3A (Cytochrome P450 3A) function was assessed using the P450-GloCYP3A4 Assay with Luciferin-IPA (Promega, Madison, WI) by adding 2 mL of the luminogenic substrate (1:1000 in WEM maintenance medium) to each liver MPS and incubated at 37°C for 1 hour with self-circulating flow in the upwards direction. Tissue formation was assessed by staining for cell nuclei on scaffolds using Hoescht 33342 (Thermo Fisher, Waltham MA) diluted 1:1000 in WEM for 20 minutes. Following image analysis, scaffolds were incubated in RIPA buffer at 4°C then scraped to remove cells; total protein was measured using a BCA ELISA kit (Pierce). Human serum albumin (HSA) levels were normalized to the total amount of protein present.