Neurovascular Organotypic Culture Models Using Induced Pluripotent Stem Cells to Assess Adverse Chemical Exposure Outcomes

Abstract

Introduction: Human-induced pluripotent stem cells (iPSCs) represent a promising cell source for the construction of organotypic culture models for chemical toxicity screening and characterization. Materials and Methods: To characterize the effects of chemical exposure on the human neurovasculature, we constructed neurovascular unit (NVU) models consisting of endothelial cells (ECs) and astrocytes (ACs) derived from human-iPSCs, as well as human brain-derived pericytes (PCs). The cells were cocultured on synthetic poly(ethylene glycol) (PEG) hydrogels that guided the self-assembly of capillary-like vascular networks. High-content epifluorescence microscopy evaluated dose-dependent changes to multiple aspects of NVU morphology. Results: Cultured vascular networks underwent quantifiable morphological changes when incubated with vascular disrupting chemicals. The activity of predicted vascular disrupting chemicals from a panel of 38 compounds (U.S. Environmental Protection Agency) was ranked based on morphological features detected in the NVU model. In addition, unique morphological neurovascular disruption signatures were detected per chemical. A comparison of PEG-based NVU and Matrigel Ô-based NVU models found greater sensitivity and consistency in chemical detection by the PEG-based NVU models. Discussion: We suspect that specific morphological changes may be used for discerning adverse outcome pathways initiated by chemical exposure and rapid mechanistic characterization of chemical exposure to neurovascular function. Conclusion: The use of human stem cell-derived vascular tissue and PEG hydrogels in the construction of NVU models leads to rapid detection of adverse chemical effects on neurovascular stability. The use of multiple cell types in coculture elucidates potential mechanisms of action by chemicals applied to the model.

Induced pluripotent stem cells for cardiovascular therapeutics: Progress and perspectives

“…Also, human iPSC-derived brain microvascular ECs and neural progenitor cells have been incorporated in the dual-channel spinal cord chip system and used as an in vitro model of human vascularized motor neural tissue (Sances et al, 2018). The blood-brain barrier chip system has also been created using human iPSC-derived brain microvascular ECs, neurons, astrocytes, and brain-like pericytes, representing an in vitro neurovascular unit that provides an important platform for modeling neurological disorders and drug screening (Lippmann et al, 2014;Qian et al, 2017;Canfield et al, 2019;Jamieson et al, 2019;Li et al, 2019;Martins Gomes et al, 2019;Neal et al, 2019;Nguyen et al, 2019;Vatine et al, 2019). Cells in such systems can be generated from the same iPSCs donor source, which gives an isogenic in vitro model (Canfield et al, 2019).…”

Studying the Inflammatory Responses to Amyloid Beta Oligomers in Brain-Specific Pericyte and Endothelial Co-Culture From Human Stem Cells

“…In vitro BBB models derived from human-induced pluripotent stem cells (hiPSCs) have shown promising results as microphysiological systems to study BBB function and neurovascular unit (Campisi et al, 2018;Brown et al, 2019;Canfield et al, 2019;Nguyen et al, 2019;Browne et al, 2021). The most common BBB models would include a co-culture of two, three, or four cell types, which usually use primary brain pericytes due to the limitation in efficiently differentiating brain pericytes from hiPSCs (Lauschke et al, 2017).…”