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

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.