Absolute protein levels and enzyme activities in human hepatocytes from three donors kept in sandwich culture (ECM overlay, 0.01 μM DEX)
P450 and UGT proteins were determined from microsomal fractions, and transporter proteins were determined from plasma membrane preparations. Concentrations are calculated as the total amount of microsomal and plasma membrane protein, respectively. P450 enzyme activities were determined in situ by measuring phenacetin O-deethylation (CYP1A2), bupropion 2-hydroxylation (CYP2B6), amodiaquine N-deethylation (CYP2C8), diclofenac 4′-hydroxylation (CYP2C9), S-mephenytoin 4′-hydroxylation (CYP2C19), and testosterone-6β hydroxylation (CYP3A4).
| Target | HuHep9966 | HuHep10261 | HuHep10901 | |||
|---|---|---|---|---|---|---|
| Protein (±S.E.M.) | Activity (±S.E.M.) | Protein (±S.E.M.) | Activity (±S.E.M.) | Protein (±S.E.M.) | Activity (±S.E.M.) | |
| fmol/μg | pmol · min−1 · mio cells−1 | fmol/μg | pmol · min−1 · mio cells−1 | fmol/μg | pmol · min−1 · mio cells−1 | |
| CYP1A2 | 3.92 ± 0.31 | 38.8 ± 0.7 | 6.88 ± 0.33 | 10.9 ± 0.6 | 11.10 ± 0.26 | 61.0 ± 0.5 |
| CYP2A6 | 7.86 ± 0.24 | 2.16 ± 0.24 | 16.94 ± 0.40 | |||
| CYP2B6 | 0.00 ± 0.00 | 10.4 ± 0.5 | 0.00 ± 0.00 | 2.5 ± 0.1 | 1.06 ± 0.13 | 24.1 ± 0.6 |
| CYP2C8 | 5.30 ± 0.68 | 63.3 ± 1.6 | 1.69 ± 0.90 | 4.8 ± 0.1 | 10.93 ± 1.24 | 296.6 ± 17.4 |
| CYP2C9 | 91.00 ± 0.72 | 92.1 ± 2.0 | 58.88 ± 0.34 | 33.1 ± 1.4 | 164.00 ± 3.12 | 293.3 ± 8.4 |
| CYP2C19 | 0.00 ± 0.00 | 0.5 ± 0.0 | 1.44 ± 0.18 | 3.2 ± 0.1 | 4.06 ± 0.23 | 37.3 ± 0.9 |
| CYP2D6 | 7.76 ± 0.28 | 5.14 ± 0.25 | 9.11 ± 0.49 | |||
| CYP2E1 | 2.92 ± 0.09 | 3.14 ± 0.08 | 13.41 ± 0.38 | |||
| CYP3A4* | 11.56 ± 4.66 | 276.9 ± 12.8 | 10.41 ± 2.49 | 34.9 ± 1.0 | 17.08 ± 4.22 | 166.8 ± 1.3 |
| CYP3A43 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | |||
| CYP3A5 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.91 ± 0.00 | |||
| CYP3A7 | 1.69 ± 4.69 | 7.63 ± 2.53 | 16.83 ± 4.31 | |||
| CYP4A11 | 15.78 ± 0.48 | 9.26 ± 0.27 | 8.33 ± 0.29 | |||
| P450 reductase | 50.01 ± 1.39 | 39.94 ± 1.62 | 60.50 ± 4.14 | |||
| UGT1A1 | 16.10 ± 1.57 | 17.43 ± 0.48 | 33.53 ± 0.68 | |||
| UGT2B7 | 72.25 ± 0.58 | 82.63 ± 3.26 | 133.0 ± 3.37 | |||
| Na/K-ATPase | 9.48 ± 0.48 | 11.64 ± 0.41 | 13.86 ± 0.32 | |||
| γ-GTP | 0.94 ± 0.05 | 6.33 ± 0.26 | 6.73 ± 0.21 | |||
| ABCA8 | 0.89 ± 0.05 | 6.28 ± 0.20 | 0.81 ± 0.02 | |||
| MDR1 | 5.82 ± 0.18 | 30.50 ± 1.81 | 5.28 ± 0.26 | |||
| MDR3 | 3.95 ± 0.25 | 11.31 ± 1.26 | 4.30 ± 0.26 | |||
| BSEP | 7.87 ± 0.15 | 7.49 ± 0.38 | 5.75 ± 0.22 | |||
| MRP1 | 0.33 ± 0.03 | 3.46 ± 0.16 | 0.32 ± 0.03 | |||
| MRP2 | 7.36 ± 0.14 | 24.05 ± 1.49 | 6.11 ± 0.11 | |||
| MRP3 | 0.00 ± 0.00 | 5.47 ± 0.53 | 0.00 ± 0.00 | |||
| MRP4 | 0.00 ± 0.00 | 0.60 ± 0.04 | 0.00 ± 0.00 | |||
| MRP5 | 0.97 ± 0.19 | 2.60 ± 0.22 | 0.56 ± 0.05 | |||
| MRP6 | 7.57 ± 0.29 | 3.20 ± 0.22 | 7.95 ± 0.21 | |||
| BCRP | 3.90 ± 0.20 | 5.23 ± 0.31 | 4.32 ± 0.19 | |||
| ABCG8 | 0.73 ± 0.04 | 5.44 ± 0.33 | 1.06 ± 0.03 | |||
| NTCP | 8.53 ± 0.59 | 12.14 ± 1.09 | 7.59 ± 1.02 | |||
| OAT2 | 4.55 ± 0.18 | 6.90 ± 0.18 | 5.58 ± 0.18 | |||
| OCT1 | 9.10 ± 0.16 | 17.38 ± 0.50 | 9.84 ± 0.16 | |||
| OATP1B3 | 3.95 ± 0.38 | 4.47 ± 0.27 | 4.46 ± 0.36 | |||
| OATP2B1 | 0.00 ± 0.00 | 6.16 ± 0.63 | 2.63 ± 0.32 | |||
| OATP1B1 | 4.85 ± 0.43 | 6.83 ± 0.42 | 4.60 ± 0.54 | |||
| CNT1 | 0.45 ± 0.04 | 3.61 ± 0.14 | 0.43 ± 0.05 | |||
| ENT1 | 2.69 ± 0.10 | 10.23 ± 0.65 | 2.22 ± 0.09 | |||
| MCT1 | 4.17 ± 0.29 | 14.31 ± 3.07 | 5.79 ± 0.48 | |||
| MATE1 | 3.50 ± 0.14 | 8.52 ± 0.85 | 2.53 ± 0.12 | |||
| Na/K-ATPase | 42.88 ± 3.15 | 29.05 ± 3.50 | 37.45 ± 2.30 | |||
| γ-GTP | 4.55 ± 0.18 | 16.03 ± 0.45 | 8.39 ± 0.25 | |||
↵* CYP3A4 protein was determined indirectly by CYP3A4 = CYP3A4/3A43 − CYP3A43.