Experimental details by company (LLC-PK1-MDR1)
Lab ID | Cell Source and Passage Number | Culture Medium | Seeding Density | Culture Time | Plate Format, Material, Pore Size, Vendor Cat. No. | Monolayer Integrity Acceptance Criteria | Substrate and Conc. | Assay Buffer and Solvent Content | Volumes AP/BL | Pre- incubationa/Assay time | Plate shaking | Positive control and Conc. |
---|---|---|---|---|---|---|---|---|---|---|---|---|
cells/well | days | nM | µl | min | rpm | µM | ||||||
09 | Netherlands Cancer Institute p. 8-32 | M199, 10% FCS, 1% 10,000 IU/µg per ml pen-strep, 100 ng/ml colchicine | 150,000–180,00 | 4 | Corning no. 3399 24-well, PC, 3.0 µm | TEER >300 Ωcm2 Lucifer Yellow: <2.5 × 10−6 cm/s | 3H-Digoxin 15 | M199 <0.5% DMSO | 300/700 | 0/120 | No | GF120918 0.5 |
10 | Netherlands Cancer Institute p. 10-30 | M199, 10% FBS, 1% PSGc | 150,000 | 4 | Millipore no. PSRP010R5 24-well, PET, 1.0 µm | Texas Red: <6% transport | 3H-Digoxin 100 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 500/500 | 0/180 | No | CyclosporineA 10 |
08 | BD Gentest cat. no. 450211 p. 14-31 | M199, 7% FBS, 0.05 mg/ml gentamycin, 0.3 mg/ml L-glutamine, 0.1 mg/ml hygromycin Bb | 40,000 | 7 | BD Falcon no. 351181 24-well, PET, 1.0 µm | TEER >150 Ωcm2 Lucifer Yellow: <2% transport | 3H-Digoxin 5000 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 100/600 | 0/90 | ∼50 | Ketoconazole 25 |
11 | BD Gentest cat. no. 450211 p. 26-37 | M199, 7% FBS, 0.05 mg/ml gentamycin, 0.1 mg/ml hygromycin Bb | 100,000 | 5–7 | Corning no. 3401 12-well, PC, 0.4 µm | TEER >400 Ωcm2 Mannitol: <2.0 × 10–6 cm/s | 3H-Digoxin 20 | HBSS pH 7.4 +10 mM HEPES +25 mM Glucose <1% DMSO | 500/1000 | 0/90 | 60–90 | Verapamil 50 |