Development of a Hydrogel Macroencapsulation Device for Improved Long-Term Islet Survival Using Injection Molding and Oxygen Modeling-Aided Design

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Description
Allogeneic islet transplantation has the potential to reverse Type 1 Diabetes in patients. However, limitations such as chronic immunosuppression, islet donor numbers, and islet survival post-transplantation prevent the widespread application of allogeneic islet transplantation as the treatment of choice. Macroencapsulation

Allogeneic islet transplantation has the potential to reverse Type 1 Diabetes in patients. However, limitations such as chronic immunosuppression, islet donor numbers, and islet survival post-transplantation prevent the widespread application of allogeneic islet transplantation as the treatment of choice. Macroencapsulation devices have been widely used in allogeneic islet transplantation due to their capability to shield transplanted cells from the immune system as well as provide a supportive environment for cell viability, but macroencapsulation devices face oxygen transport challenges as their geometry increases from preclinical to clinical scales. The goal of this work is to generate complex 3D hydrogel macroencapsulation devices with sufficient oxygen transport to support encapsulated cell survival and generate these devices in a way that is accessible in the clinic as well as scaled manufacturing. A 3D-printed injection mold has been developed to generate hydrogel-based cell encapsulation devices with spiral geometries. The spiral geometry of the macroencapsulation device facilitates greater oxygen transport throughout the whole device resulting in improved islet function in vivo in a syngeneic rat model. A computational model of the oxygen concentration within macroencapsulation devices, validated by in vitro analysis, predicts that cells and islets maintain a greater viability and function in the spiral macroencapsulation device. To further validate the computational model, pO2 Reporter Composite Hydrogels (PORCH) are engineered to enable spatiotemporal measurement of oxygen tension within macroencapsulation devices using the Proton Imaging of Siloxanes to map Tissue Oxygenation Levels (PISTOL) magnetic resonance imaging approach. Overall, a macroencapsulation device geometry designed via computational modeling of device oxygen gradients and validated with magnetic resonance (MR) oximetry imaging enhances islet function and survival for islet transplantation.
Date Created
2023
Agent

Analyzing the Survival of Hydrogel-Encapsulated Pancreatic Cells for Use in Islet Transplantation

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Description

Type 1 diabetes is a metabolic disorder in which the pancreas produces little to no insulin due to the cells being destroyed by a person’s own body. A potential treatment for this disorder is the allogeneic transplantation of pancreatic beta

Type 1 diabetes is a metabolic disorder in which the pancreas produces little to no insulin due to the cells being destroyed by a person’s own body. A potential treatment for this disorder is the allogeneic transplantation of pancreatic beta cells. Unfortunately, this potential solution requires the use of immunosuppressants. For my project with the Weaver Lab, I will be assessing pseudoislet survival in macroencapsulation via injection molding. I will be analyzing survival and metabolic assays of the pseudoislets in the mold process. Pseudoislets in hydrogels usually undergo hypoxia-included cell death due to the diffusion distances oxygen has to travel. We will test the impact of macroencapsulation device geometry on hypoxia within encapsulated cells. I will be culturing pancreatic cells and encapsulating them in hydrogels. Macroencapsulation devices will be utilized to shield islets from the immune system and eliminate the need for immunosuppressants. In order to analyze the cells’ structure and to ensure their viability, confocal microscopy will be used. Staining for live cells will be done using calcein AM which produces green fluorescence and indicates live cells. Staining for dead cells on the other hand will be done using an ethidium homodimer which produces red fluorescence and indicates dead cells. To determine if the cells are metabolically active the Alamar Blue assay will be used.

Date Created
2022-05
Agent