High Content Analysis of Human Pluripotent Stem Cell Derived Hepatocytes Reveals Drug Induced Steatosis and Phospholipidosis

Table 3

Cytotoxic effects of the tested compounds in hiPSC derived hepatocytes (hiPS-HEP) and HepG2: minimal effective concentration and toxicity risk steatotic risk index.

Cell system	Compound	Fixed assay									Live assay							TR	SRI
		V	NC		S			PL			V	NC		MMP	ROS	PMP			Lipid	ROS
		CC	MI	A	C	MI	A	C	MI	A	CC	MI	A	MI	MI	MI
hiPS-HEP	AMD	39.5	39.5	—	39.5	125	39.5	12.5	12.5	12.5	39.5	39.5	—	39.5	39.5	125	2.2	5.7	18	18
	DOX	100	—	316	316	1000	—	—	—	—	1000	100	100	1000	—	31.6	8.8	3.6	36	>114
	TET	1000	—	316	316	1000	—	—	—	—	1000	—	316	—	316	—	14.2	22.3	22	22
	SC	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	NA	NA	NA	NA
HepG2	AMD	4	39.5	125	39.5	39.5	125	—	—	—	12.5	39.5	39.5	12.5	12.5	125	2.2	1.8	18	6
	DOX	100	1000	1000	316	1000	—	—	—	—	316	—	—	1000	316	1000	8.8	11.4	36	36
	TET	316	100	316	100	100	—	—	100	—	—	1000	316	—	1000	1000	14.2	7.0	7	70
	SC	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—	NA	NA	NA	NA

Mechanism affected at the lowest concentration is denoted in bold. Statistical significance was performed using one-way ANOVA followed by Dunnett’s test. AMD: amiodarone HCl; DOX: doxycycline; TET: tetracycline; V: viability; NC: nuclear changes; S: steatosis; PL: phospholipidosis; MMP: mitochondrial membrane potential; ROS: reactive oxygen species; PMP: plasma membrane permeability; CC: cell count; MI: mean intensity; A: area; C: droplet count. Maximum plasma concentration of the drug (), The toxicity risk (TR) for each compound was defined as ratio of minimal effective concentration to the maximum plasma concentration of the drug (). Steatotic risk index (SRI) was calculated as the ratio of the MEC for neutral lipid accumulation or for ROS generation to the .