Human pluripotent stem cell-based models for predictive developmental neural toxicity
AssigneeWisconsin Alumni Research Foundation
InventorsJames A. Thomson, William L. Murphy, Charles D. Page, Michael P. Schwartz, Zhonggang Hou
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Abstract
The present invention relates to three-dimensional (3D) tissue constructs and methods of using such 3D tissue constructs to screen for neurotoxic agents. In particular, provided herein are methods of producing and using complex, highly uniform human tissue models comprising physiologically relevant human cells, where the tissue models have the degree of sample uniformity and reproducibility required for use in quantitative high-throughput screening applications.
Claims16
A method of producing a chemically defined, xenogen-free vascularized neural tissue construct, comprising:
- (a) seeding a xenogen-free, three-dimensional (3D), RGD-containing peptide-functionalized, matrix metalloproteinase (MMP)-degradable, porous hydrogel with human pluripotent stem cell-derived neural progenitor cells;
- (b) culturing the seeded hydrogel under chemically defined, xenogen-free conditions for a length of time sufficient to detect differentiation of at least a portion of the neural progenitor cells;
- (c) dispersing, on or within the cultured seeded hydrogel, human endothelial cells and human mesenchymal cells;
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- (d) culturing the seeded hydrogel comprising the dispersed human endothelial cells and human mesenchymal cells under chemically defined, xenogen-free culture conditions that promote cell differentiation, whereby a 3D chemically defined, xenogen-free, vascularized neural tissue construct comprising stratified layers of human neurons and glial cells is produced within about 14 days from seeding step (a).
Dependencies are inferred from the claim text.