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Optimizing Tolerogenic Fibroblastic Reticular Cell Seeding in Macroporous Gelatin Scaffolds for Localized Immunomodulation in Pancreatic Islet Transplantation for Type 1 Diabetes
Location: 95
Mentor: Dr. Alice Tomei
Type 1 diabetes (T1D), an autoimmune disease characterized by the destruction of insulin-producing beta cells, may be cured if pancreatic islets are successfully transplanted and protected from recurrence of autoimmunity and immune rejection. This can be achieved by co-transplanting islets with fibroblastic reticular cells (FRCs), specialized lymph node stromal cells that create supportive networks and regulate immune responses during inflammation. FRCs are candidates for providing localized immune modulation by co-transplantation with islets because they dampen inflammatory immune responses against transplanted islets. To enable this approach, we developed macroporous gelatin scaffolds to deliver FRCs locally in the islet transplantation site. While these scaffolds sustain FRC survival in vivo, approximately 40% of cells are lost during seeding in scaffolds. So, we aimed at enhancing FRC seeding efficiency in scaffolds in vitro to improve in vivo transplantation outcomes. Cell seeding onto scaffolds depends on scaffold thickness, cell number, and seeding methods. Current protocols involve applying 5µl of 200k murine FRCs onto dry scaffolds, followed by one hour of incubation, and resulting in a 60% seeding efficiency. To improve FRC seeding efficiency, we tested (1) seeding methods: soaked (pre-soaking scaffolds with media), flipped (applying cell suspension to both scaffold sides), and control (current protocol), (2) scaffold thickness (0.5 mm, 1 mm, 2 mm), and (3) cell density (100k to 1 million cells). We found that the soaked scaffold seeding condition increased the number of viable FRCs over 14 days culture as measured with a Real-Time ATP Cell Viability Assay. This study establishes a foundation for improving FRC seeding efficiency in scaffolds for enhancing FRC co-transplantation with islets, further advancing therapies for T1D treatment.
