TABLE 1

Experimental details by company (Caco-2)

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/BLPreincubationa/Assay timePlate shakingPositive control and Conc.
cells/welldaysnMµlminrpmµM
16ATCC p. 25-45DMEM, 10% FBS, 1% NEAA, 3 mM l-glutamine250,00015–18Corning no. 3401 12-well, PC, 0.4 µmPre-TEER and post-TEER <30% change3H-Digoxin 13HBSS pH 7.4 +25 mM HEPES + 5.56 mM Glucose <1% DMSO500/150015/45450Ketoconazole 50
25ATCC p. 23-35DMEM, 10% FBS, 1% NEAA, 1% pen-strep, 1% l-glutamine90,00021–25Corning no. 3401 12-well, PC, 0.4 µmPre-TEER and post-TEER <50% change3H-Digoxin 100HBSS pH 7.4 +10 mM HEPES <1% DMSO400/120060/60NoLY335979 5
17ATCC p. 25-45DMEM High Glucose, 10% FBS, 1% NEAA40,00021–25BD Falcon cat. no. 351181 24-well, PET, 1.0 µmTEER > 400 Ωcm2 Lucifer Yellow: <1.0 × 10−6 cm/s3H-Digoxin 1,000HBSS pH 7.4 +25 mM HEPES <0.5% DMSO400/95030/120NoKetoconazole 20
13INSERM U505 p. 18-30DMEM, 10% FBS, 1% pen-strep, 1% NEAA100,0007Corning no. 3401 12-well, PC, 0.4 µmLucifer Yellow: <1.0 × 10−6 cm/s3H-Digoxin 5000HBSS pH 7.4 +20 mM HEPES <2% DMSO800/80030/120NoKetoconazole 10
24ATCC p. 40-60DMEM High Glucose, 10% FBS, 1% NEAA, 1X l-glutamine100,00021–25Corning no. 3397 24-well, PC, 0.4 µmLucifer Yellow: <1.0 × 10−6 cm/s3H-Digoxin 50HBSS pH 7.4 +25 mM HEPES +10 mM Glucose <1% DMSO200/100030/120NoLY335979 10
12ATCC p. 47-52DMEM, 10% FBS100,00021–25BD Falcon cat. no. 351181 24-well, PET, 1.0 µmTEER > 400 Ωcm23H-Digoxin 1000HBSS pH 7.4 +10 mM HEPES <1% DMSO200/100030/120NoGF120918 4
18ATCC p. 25-35DMEM+GlutaMax, 10% FBS, 1% NEAA, pyruvate12,00018–22BD Falcon cat. no. 351131 96-well, PET, 1.0 µmTEER > 250 Ωcm2 Lucifer Yellow: <1.0 × 10-6 cm/sDigoxin 10,000HBSS pH 7.4 +10 mM HEPES <1% DMSO75/25030/120∼120 rpmLY335979 1
19ATCC p. 25-35MEM + 20% FCS, 1% NEAA, 1% pyruvate, 1% l-glutamine40,00021–28BD Falcon cat. no. 351181 24-well, PET, 1.0 µmLucifer Yellow: <1% transport3H-Digoxin 5000HBSS pH 7.4 +10 mM HEPES <1% DMSO250/10000/120150 rpmKetoconazole 20
15ATCC p. 58-76DMEM, 10% FBS, 1 mM sodium-pyruvate, 1% NEAA, 100 IU/ml penicillin, 100 µg/ml streptomycin67,20021–28Corning no. 3401 12-well, PC, 0.4 µmTEER 450–650 Ωcm2 LuciferYellow: ≤0.8 × 10−6 cm/sDigoxin 10,000HBSS pH 7.4 +10 mM HEPES +15 mM Glucose <0.8% DMSO500/150030/120NoCyclosporine A 5
14ATCC p. 20-50DMEM, 10% FBS, 1% NEAA, 1% l-glutamine, 100 IU/ml penicillin-G, 100 µg/ml streptomycin30,00018–21Corning no. 3396 24-well, PC, 0.4 µmTEER > 300 Ωcm2 Mannitol: <2.5 × 10−6 cm/s3H-Digoxin 5,000HBSS pH 7.4 +10 mM HEPES <1% DMSO200/6000/120NoKetoconazole 50
08ATCC no. HTB-37 p. 25-45DMEM, 10% FBS, 5 mM HEPES, 0.4% glucose, 1% NEAA, 50 µg/ml gentamycin40,00021–25BD Falcon cat. no. 351181 24-well, PET, 1.0 µmTEER >250 Ωcm2 Lucifer Yellow: <2% transport3H-Digoxin 5000HBSS pH 7.4 +10 mM HEPES <1% DMSO100/6000/120∼50Ketoconazole 25
  • FBS, fetal bovine serum; FCS, fetal calf serum; HBSS, Hanks’ balanced salt solution; MEM, modified Eagle’s medium; NEAA, nonessential amino acids; pen-strep, penicillin-streptomycin.

  • a Preincubation in presence of inhibitor.