Fluid shear stress modulation of hepatocyte-like cell function.
Fluid shear stress modulation of hepatocyte-like cell function.
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DOI：
10.1007/s00204-016-1689-8
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发表时间：
2016-07
影响因子：
6.1
通讯作者：
Hay DC
中科院分区：
文献类型：
--
作者：
Rashidi H;Alhaque S;Szkolnicka D;Flint O;Hay DC
新鲜分离的成人肝细胞被认为是体外研究的黄金标准工具。然而，原代肝细胞稀少、细胞周期停滞以及细胞表型的迅速丧失限制了它们的广泛应用。人胚胎干细胞和诱导多能干细胞为肝细胞样细胞提供了可再生的来源。尽管使用了各种分化方法，但HLC与原代人类肝细胞一样，在培养过程中表现出不稳定的表型。已经证明，通过添加人体生理学元素，例如细胞共培养，或者通过使用自然和/或合成表面，可以提高功能能力。在本研究中，研究了流体剪切力对HLC性能的影响。我们研究了两种重要的肝功能，细胞色素P450药物代谢和血清蛋白分泌，在静态培养和暴露于流体剪切力的情况下。我们的研究表明，流体剪切力使CYP1A2活性提高了大约五倍。与此相平行的是，对一种药物的敏感性大约增加了九倍，主要由CYP2D6代谢。除了代谢能力，流体剪切力也改善了肝细胞的表型，胎儿标志物甲胎蛋白的分泌大约减少了四倍。我们认为，这些研究强调了在基于细胞的模型中引入生理线索以改善体细胞表型的重要性。
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.