Multicellular hepatic in vitro models using NANOSTACKS(TM): human-relevant models for drug response prediction.

Drug-induced liver injury (DILI) continues to be one of the leading causes of drug attrition during clinical trials, as well as the number one cause of post-market drug withdrawal due to the limited predictive accuracy of preclinical animals and conventional in vitro models. In this study, the NANOSTACKS(TM) platform was introduced as a novel in vitro tool to build in vivo-relevant organ models for predicting drug responses. In particular, hepatic models, including monocultures of primary human hepatocytes (PHH), tricultures of PHH, human stellate cells (HSC), and human liver endothelial cells (LECs), and tetracultures of PHH, HSC, LECs, and human Kupffer cells (KC) were developed under static and orbital-induced mixing flow conditions. All hepatic models were characterised by assessing albumin, urea, CYP3A4, and ATP production. In addition, the preclinical DILI screening potential of the orbital-induced mixing flow monoculture and triculture models was assessed by testing the hepatotoxicity of zileuton, buspirone, and cyclophosphamide. NANOSTACKS™ represents a promising tool for the development of complex in vitro models.