Utility of human hepatocyte spheroids for evaluation of hepatotoxicity

Abstract

-Drug-induced hepatotoxicity is a common reason for discontinuing the development of candidate clinical drugs. In the present study, we investigated the utility of three-dimensionally cultured human hepatocytes (spheroids) for prediction of hepatotoxicity, using a panel of model drugs: acetaminophen, benzbromarone, chlorpromazine, cyclosporin A, diclofenac, fialuridine, flutamide, imipramine, isoniazid, ticlopidine and troglitazone. Cultured spheroids showed a significant increase of albumin secretion from 2 to 7 days; the secretion started to decrease at 14 days, but continued from 14 days to 21 days. The morphology of the spheroids was well maintained for 21 days. Long-term exposure of spheroids to hepatotoxic drugs resulted in concentration-dependent depression of albumin secretion and elevation of aspartate aminotransferase (AST) leakage. The estimated 50% effective concentration (IC 50 ) values for decrease of albumin secretion changed from 7 days to 14 days, but similar values were obtained at 14 and 21 days, except for diclofenac. Since the IC 50 values and the values of drug concentration inducing 1.2-fold elevation of AST leakage (F1.2) were similar at 14 and 21 days, an incubation period of 14 days was

Utility of human hepatocyte spheroids without feeder cells for evaluation of hepatotoxicity

“…4 morphology and continued secretion of albumin, which showed similar changes in Sph(f+) and Sph(f-) (Figs. 2 and 3), in accordance with our previous findings in Sph(f+) (Ogihara et al, 2015). In addition, albumin secretion from Sph(f+) without inducers were always higher than that of Sph(f-) as shown in Fig.…”

“…Sph(f+) and Sph(f-) were cultured in RM-101 medium containing the test compounds for 14 days after starting the assays (the same period as in the previous study) (Ogihara et al, 2015). The concentration of the compound was set based on our previous paper using primary 3D-cultured hepatocytes with feeder cells (Ogihara et al, 2015).…”

“…The concentration of the compound was set based on our previous paper using primary 3D-cultured hepatocytes with feeder cells (Ogihara et al, 2015). Therefore, the highest exposure dose of each test compound was set at a concentration 100 times higher than the maximum blood concentration at the clinical dose, and five exposure doses differing by a factor of three were used.…”

“…In addition, we suggested a protocol for assay of drug-metabolizing enzyme induction using these hepatocyte spheroids. Moreover, we evaluated the utility of these hepatocyte spheroids as an assay system for predicting DILI (Ogihara et al, 2015). Long-term exposure of spheroids to hepatotoxic drugs resulted in concentration-dependent reduction of albumin secretion and elevation of AST leakage.…”

“…On the other hand, other toxicity markers, such as alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and γ-glutamyl transpeptidase (γ-GTP), were not measured in the previous study (Ogihara et al, 2015), because the feeder cells used as a support to enable the hepatocytes to adhere to the substrate plate interfered with the measurement of these enzymes. Therefore, we re-investigated the utility of the spheroids for prediction of hepatotoxicity in humans using a hepatocyte lot that showed high adhesive potential even in the absence of feeder cells.…”