One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells

Abstract

Human hepatocytes display substantial functional inter-individual variation regarding drug metabolizing functions. In order to investigate if this diversity is mirrored in hepatocytes derived from different human pluripotent stem cell (hPSC) lines, we evaluated 25 hPSC lines originating from 24 different donors for hepatic differentiation and functionality. Homogenous hepatocyte cultures could be derived from all hPSC lines using one standardized differentiation procedure. To the best of our knowledge this is the first report of a standardized hepatic differentiation procedure that is generally applicable across a large panel of hPSC lines without any adaptations to individual lines. Importantly, with regard to functional aspects, such as Cytochrome P450 activities, we observed that hepatocytes derived from different hPSC lines displayed inter-individual variation characteristic for primary hepatocytes obtained from different donors, while these activities were highly reproducible between repeated experiments using the same line. Taken together, these data demonstrate the emerging possibility to compile panels of hPSC-derived hepatocytes of particular phenotypes/genotypes relevant for drug metabolism and toxicity studies. Moreover, these findings are of significance for applications within the regenerative medicine field, since our stringent differentiation procedure allows the derivation of homogenous hepatocyte cultures from multiple donors which is a prerequisite for the realization of future personalized stem cell based therapies.

Cell sources forin vitrohuman liver cell culture models

“…More evidence could be provided to show that hiPSC-derived hepatocytes can reproduce inter-individual differences in hepatic metabolism and responses to drugs. 154,155 Thus, such strategies could be helpful to recapitulate genetic polymorphisms and to determine the range of drug responsiveness in different patients before these compounds are tested in clinical trials.…”

Comparative transcriptomics of hepatic differentiation of human pluripotent stem cells and adult human liver tissue

“…Moreover, human induced pluripotent stem cells (hiPSCs) generated from the transfection of somatic cells with pluripotency factors, may be employed in drug discovery as disease models (1,4,20,31). In addition, these cells can also be utilized for patient specific treatment (1,20).…”

“…Moreover, human induced pluripotent stem cells (hiPSCs) generated from the transfection of somatic cells with pluripotency factors, may be employed in drug discovery as disease models (1,4,20,31). In addition, these cells can also be utilized for patient specific treatment (1,20). HPSCs have successfully been differentiated into hepatocytes (hPSC-HEP), sharing many features with in vivo hepatocytes, such as cytochrome P450 enzyme activity and glycogen storage, in addition to expression of several drug transporters (1,17,18,52,58).…”

“…In addition, these cells can also be utilized for patient specific treatment (1,20). HPSCs have successfully been differentiated into hepatocytes (hPSC-HEP), sharing many features with in vivo hepatocytes, such as cytochrome P450 enzyme activity and glycogen storage, in addition to expression of several drug transporters (1,17,18,52,58). Furthermore, they are applicable for long-term toxicity studies (21) and can accurately classify toxic agents (55).…”

“…The breakthrough of differentiating stem cells into hPSC-HEP with some activity of drug metabolic enzymes was accomplished by mimicking hepatogenesis in vitro (1,7,18,58).…”