Project Abstracts - New Investigators

Start Date: May 1, 2023

NIH HIRN Gateway Investigator Award Recipient 

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Abstract

Replacement of insulin-producing cells through islet transplantation in patients with type-1 diabetes (T1D) has proven to be an effective therapy, resulting in improved metabolic control and quality of life, and in prevention of long-term T1D complications. To obtain inexhaustible sources of insulin-producing cells, extensive efforts have been directed towards generating functional β cells or islet-like clusters (ILCs) from human induced pluripotent stem cells (hIPSC). However, a major barrier in generating ILCs is the considerably low yield of the stem cell differentiation process and the inability to obtain and maintain mature β cells in long-term culture. These current shortcomings in traditional culture systems motivate the development of an innovative bioengineered physiomimetic pancreatic niche. To engineer a physiologically relevant microenvironment for long-term culture of functional ILCs, we have developed an innovative approach combining three critical components: 1) the use of a through-pore microwell array to produce homogenously sized ILCs and prevent their agglomeration, 2) the incorporation of gravitational flow-based perfusion (direct/perpendicular medium flow through ILCs) to allow for removal of debris, continuous medium exchange, and provide mechanical cues, and 3) the inclusion of human pancreatic decellularized extracellular matrix (dECM) to ILC to provide more physiologically relevant biochemical cues. We propose to develop and test an open, long-term culture system for ILCs that provides robust control on the spatial, biophysical and biochemical culture microenvironment and allows for acquisition of multiple functional ILC readouts that will help optimizing those critical factors that promote the maturation and functionality of ILCs from hIPSC-derived endocrine progenitor (EN) cells. A single platform for manufacturing ILCs that incorporates these three technologies to promote differentiation, maturation, maintenance and testing of ILCs is not available. Here, we will conduct proof of concept experiments that will advance the field of islet biology and will validate an innovative platform to produce ILCs for beta cell replacement therapy in T1D. We envision that our platform can also be adopted for the long-term culture of human islets and for promoting maturation of neonatal porcine islets.