TABLE 2

Experimental details by company (LLC-PK1-MDR1)

Lab IDCell Source and Passage NumberCulture MediumSeeding DensityCulture TimePlate Format, Material, Pore Size, Vendor Cat. No.Monolayer Integrity Acceptance CriteriaSubstrate and Conc.Assay Buffer and Solvent ContentVolumes AP/BLPre- incubationa/Assay timePlate shakingPositive control and Conc.
cells/welldaysnMµlminrpmµM
09Netherlands Cancer Institute p. 8-32M199, 10% FCS, 1% 10,000 IU/µg per ml pen-strep, 100 ng/ml colchicine150,000–180,004Corning no. 3399 24-well, PC, 3.0 µmTEER >300 Ωcm2 Lucifer Yellow: <2.5 × 10−6 cm/s3H-Digoxin 15M199 <0.5% DMSO300/7000/120NoGF120918 0.5
10Netherlands Cancer Institute p. 10-30M199, 10% FBS, 1% PSGc150,0004Millipore no. PSRP010R5 24-well, PET, 1.0 µmTexas Red: <6% transport3H-Digoxin 100HBSS pH 7.4 +10 mM HEPES <1% DMSO500/5000/180NoCyclosporineA 10
08BD Gentest cat. no. 450211 p. 14-31M199, 7% FBS, 0.05 mg/ml gentamycin, 0.3 mg/ml L-glutamine, 0.1 mg/ml hygromycin Bb40,0007BD Falcon no. 351181 24-well, PET, 1.0 µmTEER >150 Ωcm2 Lucifer Yellow: <2% transport3H-Digoxin 5000HBSS pH 7.4 +10 mM HEPES <1% DMSO100/6000/90∼50Ketoconazole 25
11BD Gentest cat. no. 450211 p. 26-37M199, 7% FBS, 0.05 mg/ml gentamycin, 0.1 mg/ml hygromycin Bb100,0005–7Corning no. 3401 12-well, PC, 0.4 µmTEER >400 Ωcm2 Mannitol: <2.0 × 10–6 cm/s3H-Digoxin 20HBSS pH 7.4 +10 mM HEPES +25 mM Glucose <1% DMSO500/10000/9060–90Verapamil 50

  • For additional abbreviations, see Table 1.

  • ↵a Preincubation in presence of inhibitor.

  • ↵b Hygromycin B added to culture flasks, but not monolayers.

  • ↵c Penicillin Glutamine Streptomycin.