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

Applied In Vitro Toxicology. 2018; 
Eric H. Nguyen, Micah J. Dombroe, Debra L. Fisk, William T. Daly, Christine M. Sorenson, William L. Murphy, and Nader Sheibani
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Peptide Synthesis Six chemically defined hydrogel conditions were examined in a hydrogel array format (Fig. 1) for their ability to induce capillary-like network formation by ECs (Fig. 2A). Refer to Supplementary Methods for information on generating hydrogel formulations and constructing hydrogel arrays. Eight experimental replicates were performed per condition, but deformed or nonlevel hydrogels were excluded from analysis. Norbornene-functionalized PEG (PEGNB) molecule concentrations varied between 2.5 and 3.5 mM to control hydrogel modulus. Head-to-tail cyclized Arg-Gly-Asp-[d-Phe]-Cys (cyclic RGD; Genscript, Piscataway, NJ) and Cys-Arg-Gly-Asp-Ser (Linear RGD) peptide concentrations varied between 0.25 and 0.5 mM to control cell adhesion. Vascular endothelial growth factor (VEGF-A-165; R&D Systems, Minneapolis, MN) concentrations included in endothelial maintenance medium varied between 0, 5, and 10 ng/mL. Get A Quote

摘要

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 o... More

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