Project Abstracts - CHIB

Karen Christman, PhD, Investigator, University of California at San Diego
Steven George, MD, PhD, Investigator, University of California at Davis
Chris Hughes, PhD, Investigator, University of California at Irvine

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

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Abstract

An ideal system for identifying disease mechanisms of diabetes and screening for new therapeutics would be a renewable source of beta cells and the ability to study patient-specific cells. Such a system could help identify beta cell-intrinsic mechanisms of cell death in type I diabetes and help establish genotype-phenotype correlations. 2D cell culture systems have been the mainstay of attempts to culture human cadaveric islets or to differentiate human pluripotent cells toward the pancreatic beta cell fate. However, human islets cannot be maintained for prolonged periods of time with these systems, nor can functional beta cells be produced from hPSCs. Since current 2D culture conditions do not take into account critical cell-cell and cell- matrix interactions for beta cell development and function, there is a need for new 3D culture models of human islets that more accurately mimic the in vivo environment. Our multidisciplinary team of a pluripotent cell/islet biologist a vascular biologist and two bioengineers proposes to develop a novel in vitro platform to create a human islet micro-organ perfused with human microvessels in a microfluidic device with all components derived from a single human induced pluripotent cell source. First, we will optimize conditions and cell ratios by creating a 3D in vitro human islet micro-organ in static cultures outside the device that is comprised of islet endocrine cells, stromal cells, pancreas-specific extracellular matrix, and human endothelial cells (Aim 1). Next, we will assemble these 3D human islet micro-organs in a microfluidic device, so that nutrients are delivered and waste products are removed through a perfused capillary bed. This 3D islet micro- organ will closely mimic the dynamic metabolic changes typical for the in vivo beta cell environment (Aim 2). While a human pluripotent cell-derived islet micro-organ is the ultimate goal, we will pursue a parallel approach with each Aim, using human cadaveric islets as a cell source, as experiments with primary human islets will provide important insight into the microenvironment necessary for maintaining mature beta cells ex vivo. Our model, which fully mimics in vivo physiology and is amenable to high throughput screening, will provide a platform for identifying regulators of beta cell maturation, replication, failure, and survival and will help reveal the causes of human diabetes. Our microfluidic platform has the flexibility to combine islet micro-organs with additional micro-organs (e.g. liver) in a continuous vascular network to simulate the complex inter-organ interactions relevant to human beta cell physiology. Thus, our platform will enable studies into the role of inter-organ cross talk in the pathogenesis of diabetes.

Publications

• A vascularized 3D model of the human pancreatic islet for ex vivo study of immune cell-islet interaction
• Quantitative design strategies for fine control of oxygen in microfluidic systems
• In vivo-mimicking microfluidic perfusion culture of pancreatic islet spheroids
• Microfluidic device to attain high spatial and temporal control of oxygen
• N(6)-methyladenine DNA Modification in Glioblastoma
• Evaluation of different decellularization protocols on the generation of pancreas-derived hydrogels
• Cancer-associated Fibroblasts Support Vascular Growth through Mechanical Force
• Tissue Engineering the Vascular Tree