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.
Highlights
Static and two-dimensional (2D) culture systems have been used extensively to study human biology
hepatocyte-like cells (HLCs) were exposed to fluid shear stress (FSS), ranging from 2.9 × 10−5 to 4.7 × 10−5 dyne/cm2
Microscopic and quantitative ATP analysis revealed that BMS 827278 had a detrimental effect on the viability of HLCs (Medine et al 2013; Villarin et al 2015)
Summary
Static and two-dimensional (2D) culture systems have been used extensively to study human biology. Those systems are facile and cost-effective to use, they lack the complexity of the three-dimensional (3D) tissue microenvironment. As a result, those systems do not accurately model tissue physiology and can generate inaccurate datasets. The add-back of human physiology to cell based models is of the utmost importance. This will likely lead to an improvement in cell phenotype and more informative biological readouts from those systems (Godoy et al 2015)
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