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; Griffith, Linda G; White, Forest M.; ... 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.
Date issued
2017-03Department
Massachusetts 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 EngineeringJournal
Biomaterials
Publisher
Elsevier BV
Citation
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.
Version: Final published version
ISSN
01429612