Experimental details by company (Caco-2)
| 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 | Preincubationa/Assay time | Plate shaking | Positive control and Conc. |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| cells/well | days | nM | µl | min | rpm | µM | ||||||
| 16 | ATCC p. 25-45 | DMEM, 10% FBS, 1% NEAA, 3 mM l-glutamine | 250,000 | 15–18 | Corning no. 3401 12-well, PC, 0.4 µm | Pre-TEER and post-TEER <30% change | 3H-Digoxin 13 | HBSS pH 7.4 +25 mM HEPES + 5.56 mM Glucose <1% DMSO | 500/1500 | 15/45 | 450 | Ketoconazole 50 |
| 25 | ATCC p. 23-35 | DMEM, 10% FBS, 1% NEAA, 1% pen-strep, 1% l-glutamine | 90,000 | 21–25 | Corning no. 3401 12-well, PC, 0.4 µm | Pre-TEER and post-TEER <50% change | 3H-Digoxin 100 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 400/1200 | 60/60 | No | LY335979 5 |
| 17 | ATCC p. 25-45 | DMEM High Glucose, 10% FBS, 1% NEAA | 40,000 | 21–25 | BD Falcon cat. no. 351181 24-well, PET, 1.0 µm | TEER > 400 Ωcm2 Lucifer Yellow: <1.0 × 10−6 cm/s | 3H-Digoxin 1,000 | HBSS pH 7.4 +25 mM HEPES <0.5% DMSO | 400/950 | 30/120 | No | Ketoconazole 20 |
| 13 | INSERM U505 p. 18-30 | DMEM, 10% FBS, 1% pen-strep, 1% NEAA | 100,000 | 7 | Corning no. 3401 12-well, PC, 0.4 µm | Lucifer Yellow: <1.0 × 10−6 cm/s | 3H-Digoxin 5000 | HBSS pH 7.4 +20 mM HEPES <2% DMSO | 800/800 | 30/120 | No | Ketoconazole 10 |
| 24 | ATCC p. 40-60 | DMEM High Glucose, 10% FBS, 1% NEAA, 1X l-glutamine | 100,000 | 21–25 | Corning no. 3397 24-well, PC, 0.4 µm | Lucifer Yellow: <1.0 × 10−6 cm/s | 3H-Digoxin 50 | HBSS pH 7.4 +25 mM HEPES +10 mM Glucose <1% DMSO | 200/1000 | 30/120 | No | LY335979 10 |
| 12 | ATCC p. 47-52 | DMEM, 10% FBS | 100,000 | 21–25 | BD Falcon cat. no. 351181 24-well, PET, 1.0 µm | TEER > 400 Ωcm2 | 3H-Digoxin 1000 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 200/1000 | 30/120 | No | GF120918 4 |
| 18 | ATCC p. 25-35 | DMEM+GlutaMax, 10% FBS, 1% NEAA, pyruvate | 12,000 | 18–22 | BD Falcon cat. no. 351131 96-well, PET, 1.0 µm | TEER > 250 Ωcm2 Lucifer Yellow: <1.0 × 10-6 cm/s | Digoxin 10,000 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 75/250 | 30/120 | ∼120 rpm | LY335979 1 |
| 19 | ATCC p. 25-35 | MEM + 20% FCS, 1% NEAA, 1% pyruvate, 1% l-glutamine | 40,000 | 21–28 | BD Falcon cat. no. 351181 24-well, PET, 1.0 µm | Lucifer Yellow: <1% transport | 3H-Digoxin 5000 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 250/1000 | 0/120 | 150 rpm | Ketoconazole 20 |
| 15 | ATCC p. 58-76 | DMEM, 10% FBS, 1 mM sodium-pyruvate, 1% NEAA, 100 IU/ml penicillin, 100 µg/ml streptomycin | 67,200 | 21–28 | Corning no. 3401 12-well, PC, 0.4 µm | TEER 450–650 Ωcm2 LuciferYellow: ≤0.8 × 10−6 cm/s | Digoxin 10,000 | HBSS pH 7.4 +10 mM HEPES +15 mM Glucose <0.8% DMSO | 500/1500 | 30/120 | No | Cyclosporine A 5 |
| 14 | ATCC p. 20-50 | DMEM, 10% FBS, 1% NEAA, 1% l-glutamine, 100 IU/ml penicillin-G, 100 µg/ml streptomycin | 30,000 | 18–21 | Corning no. 3396 24-well, PC, 0.4 µm | TEER > 300 Ωcm2 Mannitol: <2.5 × 10−6 cm/s | 3H-Digoxin 5,000 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 200/600 | 0/120 | No | Ketoconazole 50 |
| 08 | ATCC no. HTB-37 p. 25-45 | DMEM, 10% FBS, 5 mM HEPES, 0.4% glucose, 1% NEAA, 50 µg/ml gentamycin | 40,000 | 21–25 | BD Falcon cat. no. 351181 24-well, PET, 1.0 µm | TEER >250 Ωcm2 Lucifer Yellow: <2% transport | 3H-Digoxin 5000 | HBSS pH 7.4 +10 mM HEPES <1% DMSO | 100/600 | 0/120 | ∼50 | Ketoconazole 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.