Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation

Abstract

Efficient protocols to differentiate human pluripotent stem cells to various tissues in combination with -omics technologies opened up new horizons for in vitro toxicity testing of potential drugs. To provide a solid scientific basis for such assays, it will be important to gain quantitative information on the time course of development and on the underlying regulatory mechanisms by systems biology approaches. Two assays have therefore been tuned here for these requirements. In the UKK test system, human embryonic stem cells (hESC) (or other pluripotent cells) are left to spontaneously differentiate for 14 days in embryoid bodies, to allow generation of cells of all three germ layers. This system recapitulates key steps of early human embryonic development, and it can predict human-specific early embryonic toxicity/teratogenicity, if cells are exposed to chemicals during differentiation. The UKN1 test system is based on hESC differentiating to a population of neuroectodermal progenitor (NEP) cells for 6 days. This system recapitulates early neural development and predicts early developmental neurotoxicity and epigenetic changes triggered by chemicals. Both systems, in combination with transcriptome microarray studies, are suitable for identifying toxicity biomarkers. Moreover, they may be used in combination to generate input data for systems biology analysis. These test systems have advantages over the traditional toxicological studies requiring large amounts of animals. The test systems may contribute to a reduction of the costs for drug development and chemical safety evaluation. Their combination sheds light especially on compounds that may influence neurodevelopment specifically. Video LinkThe video component of this article can be found at

Design of a high-throughput human neural crest cell migration assay to indicate potential developmental toxicants_suppl

“…For instance, assays have been developed that assess changes in early neural differentiation Pennings et al, 2012;Theunissen et al, 2012Theunissen et al, , 2013Krug et al, 2013b;Balmer et al, 2014;Waldmann et al, 2014;Rempel et al, 2015;Shinde et al, 2015), neurite outgrowth (Stiegler et al, 2011;Krug et al, 2013a), synaptogenesis (Harrill et al, 2011), gliogenesis (Fritsche et al, 2005) or myelination (Zurich et al, 2000(Zurich et al, , 2002.…”

Comparison of a teratogenic transcriptome-based predictive test based on human embryonic versus inducible pluripotent stem cells

“…Both in vitro hESC-based systems developed by the University of Konstanz (UKN) and Universitätsklinikum Köln (UKK) recapitulate the critical phases of embryonic development, during which cells can be exposed to various test compounds [11]. These systems have already been applied in numerous studies to identify and characterize developmental toxicants [12–15]. …”

Stem cells in toxicological research

“…This field of research is of high interest, since it offers the perspective of an unlimited supply of all relevant human cell types that can be applied for in vitro toxicity tests (Leist et al 2017). Research efforts have focused particularly on stem cell-derived cardiomyocyte and hepatocyte like cells and on developmental toxicity as well as developmental neurotoxicity (Waldmann et al 2017;Shinde et al 2015Shinde et al , 2016Meganathan et al 2015). Standardized protocols are available that allow the generation of beating cardiomyocytes derived from embryonic or induced pluripotent stem cells (Sirenko et al 2017).…”

“…For developmental toxicity testing, stem cells are allowed to differentiate to neuronal precursor cells (Shinde et al 2015;Colaianna et al 2017). Chemicals are added during the differentiation period to test whether this process is disturbed (Pallocca et al 2016;Rempel et al 2015).…”