Developing High-Fidelity Hepatotoxicity Models From Pluripotent Stem Cells

Abstract

Faithfully recapitulating human physiology "in a dish" from a renewable source remains a holy grail for medicine and pharma. Many procedures have been described that, to a limited extent, exhibit human tissue-specific function in vitro. In particular, incomplete cellular differentiation and/or the loss of cell phenotype postdifferentiation play a major part in this void. We have developed an interdisciplinary approach to address this problem, using skill sets in cell biology, materials chemistry, and pharmacology. Pluripotent stem cells were differentiated to hepatocytes before being replated onto a synthetic surface. Our approach yielded metabolically active hepatocyte populations that displayed stable function for more than 2 weeks in vitro. Although metabolic activity was an important indication of cell utility, the accurate prediction of cellular toxicity in response to specific pharmacological compounds represented our goal. Therefore, detailed analysis of hepatocellular toxicity was performed in response to a custom-built and well-defined compound set and compared with primary human hepatocytes. Importantly, stem cell-derived hepatocytes displayed equivalence to primary human material. Moreover, we demonstrated that our approach was capable of modeling metabolic differences observed in the population. In conclusion, we report that pluripotent stem cell-derived hepatocytes will model toxicity predictably and in a manner comparable to current gold standard assays, representing a major advance in the field. STEM CELLS TRANSLATIONAL MEDICINE 2013;2:505-509

Accurate Prediction of Drug-Induced Liver Injury Using Stem Cell-Derived Populations

“…Cell Culture and Differentiation hESCs (H9) and iPSCs (33D6) were cultured as described [16,[18][19][20] and maintained in a humidified 37°C, 5% CO 2 incubator. At day 9, differentiating cells from hESCs and iPSCs were cultured in hepatocyte maturation medium HepatoZYME (Life Technologies, Carlsbad, CA, http://www.lifetech.com) containing 1% Glutamax (Life Technologies), supplemented with 10 mM hydrocortisone 21-hemisuccinate (Sigma-Aldrich, St. Louis, MO, http://www.…”

“…Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are examples of renewable cell populations [9,10]. Both populations have been shown to efficiently differentiate to hepatocytes by either spontaneous or directed differentiation and, therefore, offer significant promise in the quest to deliver a new wave of predictive human models [11][12][13][14][15][16][17][18][19]. In this vein, we have developed highly efficient, scalable, and serum-free differentiation procedures that are compatible with pluripotent stem cells.…”

Prediction of interindividual differences in hepatic functions and drug sensitivity by using human iPS-derived hepatocytes

“…Human induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (HLCs) are expected to be useful for the prediction of DILI in the early phase of drug development. Many groups, including our own, have reported that the human iPS-HLCs have the ability to metabolize drugs, and thus these cells could be used to detect the cytotoxicity of drugs that are known to cause DILI (1,2). However, to accurately predict DILI, it will be necessary to establish a panel of human iPS-HLCs that better represents the genetic variation of the human population because there are large interindividual differences in the drug metabolism capacity and drug responsiveness of hepatocytes (3).…”

Highlight report: perspectives in stem cell research—unbiased quantification of the similarity between in vitro generated and primary hepatocytes

“…However, it is discussed controversially to which degree stem cell-derived cells resemble primary cells. In the field of stem cell-derived hepatocyte-like cells, some authors reported that generation of 'functional,' 'high fidelity' hepatocytes from stem and precursor cells is already possible (Medine et al 2013;Szkolnicka et al 2014;Huang et al 2014), while others present a more critical point of view Hengstler et al 2005;Brulport et al 2007).…”