Human pluripotent stem cell-derived neural constructs for predicting neural toxicity

Abstract

Human pluripotent stem cell-based in vitro models that reflect human physiology have the potential to reduce the number of drug failures in clinical trials and offer a cost-effective approach for assessing chemical safety. Here, human embryonic stem (ES) cell-derived neural progenitor cells, endothelial cells, mesenchymal stem cells, and microglia/ macrophage precursors were combined on chemically defined polyethylene glycol hydrogels and cultured in serum-free medium to model cellular interactions within the developing brain. The precursors self-assembled into 3D neural constructs with diverse neuronal and glial populations, interconnected vascular networks, and ramified microglia. Replicate constructs were reproducible by RNA sequencing (RNA-Seq) and expressed neurogenesis, vasculature development, and microglia genes. Linear support vector machines were used to construct a predictive model from RNA-Seq data for 240 neural constructs treated with 34 toxic and 26 nontoxic chemicals. The predictive model was evaluated using two standard hold-out testing methods: a nearly unbiased leave-one-out cross-validation for the 60 training compounds and an unbiased blinded trial using a single holdout set of 10 additional chemicals. The linear support vector produced an estimate for future data of 0.91 in the cross-validation experiment and correctly classified 9 of 10 chemicals in the blinded trial.organoid | machine learning | tissue engineering | differentiation | toxicology T here is a pressing need for improved methods to assess the safety of drugs and other compounds (1-5). Success rates for drug approval are declining despite higher research and development spending (6), and clinical trials often fail due to toxicities that were not identified through animal testing (7). In addition, most of the chemicals in commerce have not been rigorously assessed for safety despite growing concerns over the potential impact of industrial and environmental exposures on human health (2-5). Animal models are costly, time consuming, and fail to recapitulate many aspects of human physiology, which has motivated agencies such as the National Institutes of Health (NIH) and the US Environmental Protection Agency (EPA) to initiate programs that emphasize human cellular approaches for assessing the safety of drugs (1) and environmental chemicals (2, 3). In vitro cellular models that accurately reflect human physiology have the potential to improve the prediction of drug toxicity early in the development pipeline (1) and would provide a cost-effective approach for testing other sources of chemical exposure, including food additives, cosmetics, pesticides, and industrial chemicals (2-5).The human brain is particularly sensitive to toxic insults during development and early childhood (8), and there is growing concern that exposure to environmental chemicals may be linked to the rising incidence of neurodevelopmental disorders worldwide (4). Human brain development is mediated by highly coordinated cellular interactions between functionally ...

Functional characterization of human pluripotent stem cell-derived arterial endothelial cells

“…Previously, we developed an efficient endothelial cell differentiation protocol (15), but the resulting cells lacked strong expression of arterial markers (SI Appendix, Fig. S1 A and B).…”

Developments in 3D neural cell culture models: the future of neurotherapeutics testing?

“…Tissueengineered structures are typically made from soft and porous matrices to allow for sufficient passage and supply of oxygen and nutrients to the cells. Synthetic materials such as polycaprolactone, polyethylene glycol, and even polystyrene have been used as scaffolds for cell adherence and support [5,19,20]. Another method for scaffold-based neural cell culture provides cells with natural ECM proteins such as laminin or collagen [3,21,22].…”

The Uses of Stem Cells in Toxicology Studies

“…Schwartz ve ark. (2015) hESC'lerden sağlanan nöral öncü hücreler, endotelyal hücreler, mezenşimal kök hücreler ve mikroglia/makrofaj hücrelerini kullanarak doku mühendisliği ile in vitro nörotoksisite taramalarında faydalanılmak üzere 3 boyutlu beyin modeli geliştirmiştir [18].…”