Xenobiotic-Induced Fetal Hepatocyte Maturation

Abstract

Introduction:

Human fetal liver progenitor cells (hFLPCs) offer an emerging limitless source for generating mature hepatocytes that can be useful in cell therapies, as well as in pharmacological and toxicological studies. However, hFLPCs have very limited use over adult hepatocytes because of lower expression of drug-metabolizing enzymes. Here, we studied a novel method to achieve physiological maturity of fetal hepatocytes by exposing them to PXR and HNF4α agonist.

Materials and Methods:

We investigated the expression of cytochrome P450s (CYP3A4 and CYP2B10), glutathione S-transferase Mu 1 (GSTM1), UDP-glucuronosyltransferase 1-1 (UGT1A1), and drug transporters ATP-binding cassette subfamily C member (ABCC2 and ABCC3) by exposing them to rifampicin (Rif, 10 and 30 μM), linoleic acid (50 and 100 μM), and 0.1% dimethyl sulfoxide (DMSO) for 4, 8, and 12 days. The real-time polymerase chain reaction and western blotting were used to determine mRNA and protein expression of CYPs.

Results:

Increased CYP expression on the mRNA and protein level was detected in hFLPCs, which are exposed to DMSO- and Rif-treated cells that were much higher than in untreated. There was increase in levels of CYP2B10 protein with respect to time when hFLPCs exposed to DMSO.

Discussion and Conclusions:

Thus, this is the first report on expression of few phase I, II, and III enzymes in hFLPCs challenged with PXR and HNF4α agonists and to generate hFLPCs expressing drug-metabolizing enzymes similar to that of primary human adult hepatocytes.