Acetaminophen cytotoxicity is ameliorated in a human liver organotypic co-culture model

Abstract

Organotypic liver culture models for hepatotoxicity studies that mimic in vivo hepatic functionality could help facilitate improved strategies for early safety risk assessment during drug development. Interspecies differences in drug sensitivity and mechanistic profiles, low predictive capacity, and limitations of conventional monocultures of human hepatocytes, with high attrition rates remain major challenges. Herein, we show stable, cell-type specific phenotype/cellular polarity with differentiated functionality in human hepatocyte-like C3A cells (enhanced CYP3A4 activity/albumin synthesis) when in co-culture with human vascular endothelial cells (HUVECs), thus demonstrating biocompatibility and relevance for evaluating drug metabolism and toxicity. In agreement with in vivo studies, acetaminophen (APAP) toxicity was most profound in HUVEC mono-cultures; whilst in C3A:HUVEC co-culture, cells were less susceptible to the toxic effects of APAP, including parameters of oxidative stress and ATP depletion, altered redox homeostasis, and impaired respiration. This resistance to APAP is also observed in a primary human hepatocyte (PHH) based co-culture model, suggesting bidirectional communication/stabilization between different cell types. This simple and easy-to-implement human co-culture model may represent a sustainable and physiologically-relevant alternative cell system to PHHs, complementary to animal testing, for initial hepatotoxicity screening or mechanistic studies of candidate compounds differentially targeting hepatocytes and endothelial cells.

Human Stem Cell-Derived Endothelial-Hepatic Platform for Efficacy Testing of Vascular-Protective Metabolites from Nutraceuticals

“…Conversely, we also investigated whether hPSC‐ECs could, in turn, impact the metabolic function of hPSC‐HEPs in a coculture setting. It has been reported that endothelial cells could improve hepatic function [] and provide some levels of hepatoprotection from acetaminophen toxicity []. In our study, cocultured hPSC‐HEPs had comparable albumin levels with monocultured hPSC‐HEPs (supplemental online Fig.…”

“…Multicellular coculture models [, , ] are gaining momentum for various applications. A recent study described the use of human umbilical vein endothelial cells to stabilize hepatoblastoma C3A cells and modulate drug‐induced hepatotoxicity []. Endothelial cells from hPSCs were used to vascularize liver constructs [].…”

“…Endothelial cells from hPSCs were used to vascularize liver constructs []. These are part of the advances to model liver vasculatures, using multiple stromal cell types alongside hepatocytes to develop liver lobule and sinusoid‐like structures [, , ], for hepatotoxicity screening and tissue‐engineering applications. We are the first to report a human stem cell‐derived endothelial‐hepatic platform for efficacy testing of complex compounds.…”

Activation of Wnt/β-catenin signalling via GSK3 inhibitors direct differentiation of human adipose stem cells into functional hepatocytes

“…Similarly, the levels of CYP2C9 and CYP3A4 enzyme activity in hASC-HLCs-Chir99021 were relatively higher than that in hASC-HLCs-activin A. These data imply that hASC-HLCs in this study more closely resemble human hepatocytes for pre-clinical assessment of new candidate compounds in drug development33. Of course many efforts are still needed for promoting hASC-HLCs further maturation under the hepatic microenvironment, until they have equal metabolic abilities compared with human adult hepatocytes.…”

Type of endothelial cells affects HepaRG cell acetaminophen metabolism in both 2D and 3D porous scaffold cultures

“…It is not clear what interactions caused decreased viability of HepaRG cells in the presence of HUVECs relative to LSECs. In addition, these interactions seem to be unique as the results obtained using the HepG2‐derived C3A cell line, in combination with HUVEC is shown to promote the survival of hepatocytes (Nelson et al, ). Those co‐cultures showed improvements in albumin synthesis by day 3 and less toxicity towards APAP.…”