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. Author manuscript; available in PMC: 2023 Nov 5.
Published in final edited form as: Comput Toxicol. 2022 Nov 5;24:10.1016/j.comtox.2022.100250. doi: 10.1016/j.comtox.2022.100250

Table 2:

in vitro toxicity and toxicokinetic testing

Toxicological
and
Toxicokinetic
Responses
Assay Assay
Endpoints
Purpose Assay
outcomes
Availability/References
Developmental Toxicity Zebrafish embryo assay Developmental progression Assess potential teratogenicity Zebrafish point of departure (POD) and development information Internal EPA assay
doi:10.1016/j.reprotox.2011.10.018
Immunotoxicity Bioseek Diversity Plus Protein biomarkers across multiple primary cell types Measure potential disease and immune responses 148 individual assays which report hitcall and AC50 Commercial
doi:10.1177/1087057109345;
doi:10.1038/nbt.2914
Developmental Neurotoxicity Microelectrode array assay (rat primary neurons) Neuronal electrical activity Impacts on development of functional neural networks 17 different parameters measured to culminate in one in vitro POD Internal EPA assay
doi:10.1007/s00204-017-203505
Endocrine Disruption ACEA real-time cell proliferation assay (T47D), plus selected endpoints from Attagene Estrogen-dependent cell proliferation Functional measurement of estrogen receptor activity 2 individual assays which report hitcall and AC50 Commercial
doi:10.1021/tx400117y;
doi:10.1016/j.tiv.2005.12.008
General Toxicity / Bioactivity Attagene cis- and trans-Factorial assays Nuclear receptor and other transcription factor activation in HepG2 cells Activation of transcription factors involved in critical cell pathways and stress responses 81 individual assays which report hitcall and AC50 Commercial
doi:10.1038/nmeh.1186;
doi:10.1021/tx900325g
High-throughput transcriptomic assay Cellular mRNA, 2D HepaRG and U2OS cells Measures changes in important biological pathways 1000s of genes which are used to derive an in vitro POD Internal EPA assay with contract support
doi:10.1371/journal.pone.0178302
High-throughput phenotypic profiling Nuclear, endoplasmic reticulum, nucleoli, golgi, plasma membrane, cytoskeleton, and mitochondria morphology, U2OS and MCF7 cells Changes in cellular organelles and general morphology Cell painting (CP) and cell viability (CV) PODs. 1300 features aggregated into 49 categories which are used to derive the CP POD. Internal EPA assay
doi:10.1177/2472555220928004;
doi:10.1016/j.taap.2019.114876;
doi:10.1038/nprot.2016.105
Intrinsic hepatic clearance Hepatocyte stability assays Time course metabolism of parent chemical in primary human hepatocytes Measure metabolic breakdown by the liver Clearance (Cl) Commercial and Internal EPA and NTP assay
doi:10.1124/dmd.30.8.892
Plasma protein binding Ultra centrifugation assay Fraction of chemical not bound to plasma protein Measure amount of free chemical in the blood Fraction unbound Commercial and Internal EPA assay
doi:10.1002/jps.21317