On-demand dissolution of modular, synthetic extracellular matrix reveals local epithelial-stromal communication networks
Author(s)Ahrens, Caroline Chopko; Valdez Macias, Jorge Luis; Cook, Christi Dionne; Wang, Alex J-S; Brown, Alexander Thomas; Kumar, Manu Prajapati; Stockdale, Linda; Rothenberg, Daniel Abram; Renggli-Frey, Kasper; Gordon, Elizabeth A; Lauffenburger, Douglas A; White, Forest M; Griffith, Linda G; ... Show more Show less
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Methods to parse paracrine epithelial-stromal communication networks are a vital need in drug development, as disruption of these networks underlies diseases ranging from cancer to endometriosis. Here, we describe a modular, synthetic, and dissolvable extracellular matrix (MSD-ECM) hydrogel that fosters functional 3D epithelial-stromal co-culture, and that can be dissolved on-demand to recover cells and paracrine signaling proteins intact for subsequent analysis. Specifically, synthetic polymer hydrogels, modified with cell-interacting adhesion motifs and crosslinked with peptides that include a substrate for cell-mediated proteolytic remodeling, can be rapidly dissolved by an engineered version of the microbial transpeptidase Sortase A (SrtA) if the crosslinking peptide includes a SrtA substrate motif and a soluble second substrate. SrtA-mediated dissolution affected only 1 of 31 cytokines and growth factors assayed, whereas standard protease degradation methods destroyed about half of these same molecules. Using co-encapsulated endometrial epithelial and stromal cells as one model system, we show that the dynamic cytokine and growth factor response of co-cultures to an inflammatory cue is richer and more nuanced when measured from SrtA-dissolved gel microenvironments than from the culture supernate. This system employs accessible, reproducible reagents and facile protocols; hence, has potential as a tool in identifying and validating therapeutic targets in complex diseases.
DepartmentMassachusetts Institute of Technology. Biotechnology Process Engineering Center; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Chemical Engineering; Massachusetts Institute of Technology. Department of Mechanical Engineering
Valdez, Jorge, Christi D. Cook, Caroline Chopko Ahrens, Alex J. Wang, Alexander Brown, Manu Kumar, Linda Stockdale, et al. “On-Demand Dissolution of Modular, Synthetic Extracellular Matrix Reveals Local Epithelial-Stromal Communication Networks.” Biomaterials 130 (June 2017): 90–103.
Final published version