Brain MAPs

Project Summary

The Brain MAPs project objective is to generate a high throughput neurotoxicity screening platform that recapitulates the phenotypic diversity within the human central nervous system (hCNS) while still being sensitive enough to detect toxicity towards a single phenotype. Novel differentiation protocols and innovative microfabrication techniques, using the HMAP center Microscale System Core, will be used to generate a microfluidic screening platform comprised of an array of 3D organotypic tissues derived from human pluripotent stem cells and differentiated into region-specific tissues that span the hCNS. Collectively, we hypothesize that the tissues will capture an unparalleled amount of hCNS phenotypic diversity, and their development will be modeled using RNA-seq analysis to comprehensively capture this diversity and provide fundamental insights into signaling pathways that govern their development, function, and can potentially be targeted by toxins. Furthermore, we will use Cas9-mediated genome editing to increase the versatility of our organotypic hCNS model. In combination with high-content imaging, our microfluidic screening platform will be ideal for executing quantitative high throughput screening assays to generate predictive dose-response data on toxins with neural phenotype-specific effects.

Brain MAPs diagram
Schematic of (A) rostrocaudal and (B) dorsoventral domains that will be included in the CNS model. Standardized protocols have been developed for domains encircled in green. (C) Score plot of PCA on time course hindbrain patterning study with K-means clustering indicating 5 distinct transcriptional profiles. (D) Heat map of QPCR results from dorsoventral patterning of cervical spinal cord tissues. (E) Schematic of microfluidic platform (TOP) with cells seeded between media reservoirs (MIDDLE) containing soluble factors, 300μm scale bar. Neurally differentiating hPSCs display normal N-cadherin polarization behavior (BOTTOM) within the platform.

Prof. Randolph Ashton

Brain MAPs
Principal Investigator

Room 4165,
330 North Orchard Street,
Madison WI 53715

Our goal is to understand, model, and recapitulate in vitro the instructive signals utilized by human embryos to pattern tissue-specific differentiation of pluripotent stem cells, and apply this knowledge towards the rational design of tissue engineered scaffolds and other regenerative therapeutic strategies. Currently, we primarily focus on generating tissues and therapies for the central nervous system.