Bio-MAPS: BioMolecular-Array Patterns for Precision Differentiation of Intestinal Stem Cells

Controlling cell differentiation is critical when manufacturing products for tissue engineering and regenerative medicine. In the body, cell differentiation is governed by patterns of biochemical and mechanical features in tissues. Numerous techniques have been proposed to mimic these patterns, but their precision and reproducibility need improvement.

In this effort, the research team aims to establish a new technology to build biochemical patterns directing the differentiation of intestinal stem cells, which are important for understanding gastrointestinal function and disease. Differentiated cells will be harvested and used as building blocks to construct models of the gut wall. The results of this effort will transform our understanding of how tissue biochemistry determines cell behaviors in the intestine.

This will in turn provide a unique toolbox to formulate new “biomaterial recipes” for manufacturing cells that are “regenerative medicine-ready." To broaden impact, the technological outcomes of this research will be integrated in academic curricula, new tutorials via the Comparative Medicine Institute, and create ad hoc courses on cell production for biotech professionals. The research team will also connect with research and industrial stakeholders at NIIMBL, an institute in the Manufacturing USA network, to evaluate the use of their technology for the engineering of therapeutic cells and tissues.

Of paramount interest to tissue engineers is the precise differentiation of stem cells that are utilized as building blocks to construct organoids and microphysiological models for investigating developmental biology, pathology, and drug discovery. The project's approach to cell differentiation materials introduces Bio-Molecular-Array Patterns (Bio-MAPs), i.e., combined 3D gradients of cell signaling factors integrated within polymer matrices via ligand-mediated (non-covalent) adsorption.

This concept will be demonstrated for the precision differentiation of multi-potent intestinal stem cells. Accordingly, the project will adopt a biomolecular palette comprising collagen (COL), BMP-2, Wnt-3A, and Jagged-like ligands (which stimulate the Notch pathway), whose intersecting gradients in the intestinal crypt and villus are known to direct cell differentiation.

Single-gradient Bio-MAPs (constant collagen plus gradients of laminin, BMP2, or Wnt3) will be developed to correlate Bio-MAP architecture (i.e., concentration and gradients of combined factors) to the adhesion, proliferation, and differentiation of human ISC and subsequent lineages and to identify design rules and patterns that maintain stem-like phenotypes vs. direct differentiation towards secretory (mucus-producing) lineages or absorptive enterocytes. The project will develop complex-gradient Bio-MAPs (COL, BMP-2,Wnt-3, and Jagged-like ligands), where combinations of coherent or opposing signaling factors are utilized as backgrounds against tertiary presentation of Notch-binding ligands - herein, Jagged-like ligand (JLL) - to identify synergistic patterns that direct the differentiation towards quantitatively predictable phenotypic and genotypic outcomes.

Complementary analytical and cytological techniques will be coupled to elucidate fundamental synergistic mechanisms of the select signaling factors and their geometric display.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.