title: Fluid shear stress modulation of hepatocyte-like cell function.
abstract: Freshly isolated human adult hepatocytes are considered to be the gold standard tool for in vitro studies. However, primary hepatocyte scarcity, cell cycle arrest and the rapid loss of cell phenotype limit their widespread deployment. Human embryonic stem cells and induced pluripotent stem cells provide renewable sources of hepatocyte-like cells (HLCs). Despite the use of various differentiation methodologies, HLCs like primary human hepatocytes exhibit unstable phenotype in culture. It has been shown that the functional capacity can be improved by adding back elements of human physiology, such as cell co-culture or through the use of natural and/or synthetic surfaces. In this study, the effect of fluid shear stress on HLC performance was investigated. We studied two important liver functions, cytochrome P450 drug metabolism and serum protein secretion, in static cultures and those exposed to fluid shear stress. Our study demonstrates that fluid shear stress improved Cyp1A2 activity by approximately fivefold. This was paralleled by an approximate ninefold increase in sensitivity to a drug, primarily metabolised by Cyp2D6. In addition to metabolic capacity, fluid shear stress also improved hepatocyte phenotype with an approximate fourfold reduction in the secretion of a foetal marker, alpha-fetoprotein. We believe these studies highlight the importance of introducing physiologic cues in cell-based models to improve somatic cell phenotype.
authors.affiliation: MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, EH16 4UU, UK.
authors.forename: Hassan
authors.initials: H
authors.lastname: Rashidi
authors.affiliation: MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, EH16 4UU, UK.
authors.forename: Sharmin
authors.initials: S
authors.lastname: Alhaque
authors.affiliation: MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, EH16 4UU, UK.
authors.forename: Dagmara
authors.initials: D
authors.lastname: Szkolnicka
authors.affiliation: MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, EH16 4UU, UK.
authors.forename: Oliver
authors.initials: O
authors.lastname: Flint
authors.affiliation: MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, EH16 4UU, UK. davehay@talktalk.net.
authors.forename: David C
authors.identifier: http://orcid.org/0000-0002-7593-5973
authors.initials: DC
authors.lastname: Hay
mesh_terms: D000418:Albumins / Q000378:metabolism; D001696:Biomechanical Phenomena; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002470:Cell Survival / Q000502:physiology; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D013312:Stress, Physiological / Q000502:physiology*; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: D000418:Albumins; D000509:alpha-Fetoproteins; C504573:CYP1A2 protein, human; D019388:Cytochrome P-450 CYP1A2; D019389:Cytochrome P-450 CYP2D6
keywords: Albumin secretion; Alpha-fetoprotein secretion; Cytochrome P450 Metabolism; Embryonic stem cell; Fluid shear stress; Hepatocyte-like cell