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

Table 5

Comparison of assay imprecision for each parameter measured in hiPSC derived hepatocytes (hiPS-HEP) and HepG2.

Assay parameter	Variation between wells in a plate		Variation between plates in a batch		Variation between batches
	hiPS-HEP	HepG2	hiPS-HEP	HepG2	hiPS-HEP	HepG2
	CV	CV	CV%	CV%	CV%	CV%
CC (fixed)	6.8 ± 4.8	8.9 ± 2.6	13.0	4.1	26.7	22.6
CC (live)	15.3 ± 12.3	18.9 ± 17.3	25.8	25.5	35.3	35.0
NC
MI	2.4 ± 1.3	1.0 ± 0.6	6.0	22.3	4.1	12.9
Area	2.3 ± 1.1	2.2 ± 0.9	6.4	6.7	5.5	6.9
ST
MI	6.3 ± 2.7	5.0 ± 4.6	9.9	5.7	21.4	8.6
Area	3.9 ± 3.1	3.5 ± 4.5	1.1	10.7	6.6	9.5
PH
MI	3.2 ± 1.5	1.8 ± 1.4	7.9	4.5	8.1	4.0
Area	2.4 ± 1.0	10.2 ± 10.5	18.1	37.0	13.3	2.8
MMP
MI	4.7 ± 6.2	3.4 ± 2.7	12.3	22.1	6.8	22.4
ROS
MI	0.9 ± 0.84	4.1 ± 2.8	4.0	7.3	5.8	5.2
PMP
MI	4.8 ± 3.02	2.6 ± 2.0	3.9	14.7	0.6	23.0

Imprecisions of cell-to-cell within a well, well-to-well within a plate, plate-to-plate within a batch, and batch-to-batch are compared for all parameters measured. For calculating well-to-well variance, a mean, SD, and CV% were calculated for every control well () and an average CV% of all the eight plates was reported for well-to-well variation. The well-to-well mean values for each parameter within a batch () were then averaged and its CV% was calculated. The average CV% from the four batches was used to show plate-to-plate variation. For batch-to-batch variance, the mean values from each plate were averaged and its CV% was reported ().