Project Abstracts - CHIB

Ashutosh Agarwal, PhD, Investigator, University of Miami
Camillo Ricordi, MD, Investigator, University of Miami
Peter Buchwald, PhD, co-Investigator, University of Miami
Ed Phelps, PhD, co-Investigator, University of Miami

Start Date: September 25, 2014
End Date: June 30, 2019

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Abstract

Engineering a Human Physiomimetic Islet Microsystem Type 1 diabetes mellitus, an autoimmune disease resulting in destruction of the insulin- producing pancreatic beta cells, is one of the most common and costly chronic pediatric diseases. A significant impediment to understanding disease pathology and the development of cellular replacement therapies for Type 1 diabetes is the inability to sustain mature human beta cells in culture. In this proposal, we seek to engineer physiomimetic 3D niches within microfluidics devices for maturation, maintenance, and monitoring of human beta cells via the convergence of technologies from cell biology, matrix engineering, micro/nano fabrication, and microsensors. The microfluidic devices will connect to universal docks and provide intimate control over the cellular microenvironment by independent and simultaneous modulation of liquid and gas phases, multiparameteric monitoring, and assessment of cellular readouts and samplers for off-line biochemical analyses. With this degree of control, the effect of various niche parameters on human islet maintenance and generation of mature islets from human pancreatic precursors can be clearly delineated. Of particular interest in this application are the contributions of the physiological and extracellular matrix environment on islet health and maturation. Physiological oxygen, a critical parameter in steering pancreatic progenitor differentiation towards endocrine lineage, can be intimately modulated on the microscale via the control afforded by the microfabricated platform. Further, systematic evaluation of the contributions of matrix components on promoting islet health and directing islet differentiation within controlled 3D niches is feasible via tailored presentation of native extracellular matrix components. The ultimate goals of this proposal are twofold: 1) engineer a microfabricated “device and dock” system capable of providing microscale control of soluble and physiological conditions and agile assessment of multiple functional readouts in an enclosed, long-term culture system; and 2) utilize this innovative platform to systematically delineate critical factor capable of supporting both human islet maintenance and maturation of islet-like structures from human pancreatic progenitor cells. The project builds on recent breakthroughs by our team in creating microphysiological systems for other organ systems, engineering perifusion systems, matrix engineering, recreating oxygen controlled microenvironments, and progenitor differentiation. As such, the multidisciplinary consortium assembled herein is well poised to address these grand challenges.

Publications

• Organoid microphysiological system preserves pancreatic islet function within 3D matrix
• A Chemically Defined Common Medium for Culture of C2C12 Skeletal Muscle and Human Induced Pluripotent Stem Cell Derived Spinal Spheroids
• Nanotechnology Approaches to Modulate Immune Responses to Cell-based Therapies for Type 1 Diabetes
• Enzymatically crosslinked gelatin-laminin hydrogels for applications in neuromuscular tissue engineering
• The Effect of Recovery Warm-up Time Following Cold Storage on the Dynamic Glucose-stimulated Insulin Secretion of Isolated Human Islets
• Mechanism and effects of pulsatile GABA secretion from cytosolic pools in the human beta cell
• Concentration-Dependency and Time Profile of Insulin Secretion: Dynamic Perifusion Studies With Human and Murine Islets
• Engineering immunomodulatory biomaterials for type 1 diabetes
• 3-D physiomimetic extracellular matrix hydrogels provide a supportive microenvironment for rodent and human islet culture
• Glucose-Stimulated Insulin Release: Parallel Perifusion Studies of Free and Hydrogel Encapsulated Human Pancreatic Islets
• Metabolomics Study of the Effects of Inflammation, Hypoxia, and High Glucose on Isolated Human Pancreatic Islets
• Resealable, optically accessible, PDMS-free fluidic platform for ex vivo interrogation of pancreatic islets
• Biomimetic on-a-chip platforms for studying cancer metastasis
• Engineering biomimetic materials for islet transplantation
• Thermoresponsive release of viable microfiltrated Circulating Tumor Cells (CTCs) for precision medicine applications
• Experimental evaluation and computational modeling of the effects of encapsulation on the time-profile of glucose-stimulated insulin release of pancreatic islets