Download A Comparative Analysis of the Cellular Composition of Porcine Islets

A Comparative Analysis of the Cellular Composition of Porcine Islets and Stem Cell-Derived Insulin
Producing Cells
Miranda Stiewig, KT Ho-Nguyen
Mentors: Rahul Krishnan, Jonathan RT Lakey
Type I Diabetes is an autoimmune disorder in which the patient’s pancreatic β cells are progressively
destroyed, rendering the body unable to produce insulin, the hormone which regulates blood glucose. While
conventionally, Type I diabetes is managed with lifelong insulin administration, alternative treatments include
xenotransplantation of pancreatic islets or allotransplantation of insulin-producing cells (IPCs) derived from
human embryonic stem cells (hESCs) are being evaluated. The aim of this study was to compare the cellular
composition of juvenile porcine pancreatic islets and hESC-IPCs during in vitro culture in order to determine
which cell type expresses more insulin and is thus a better candidate for transplantation in type 1 diabetics.
We predicted that both types of cells would show increased endocrine, specifically insulin-producing, tissue
after in vitro culture. Porcine islets were also expected to show decreased exocrine tissue over prolonged in
vitro culture (up to 14 days). Juvenile porcine islets were analyzed for insulin, glucagon and amylase
expression using flow cytometry after 3, 7 and 14 days of in vitro culture. hESC-IPCs were evaluated for
insulin and glucagon expression on the day of receipt and after 5 days of in vitro culture. The hESC-IPCs
demonstrated an 8.35 ± 3.65% increase in insulin and 11.2 ±1.55% glucagon expression by Day 5. The
porcine islets demonstrated 45.27±15.14% higher insulin expression by day 14 compared to day 3, while
amylase expression decreased by 24.24±5.84%. Our study demonstrates that prolonged in vitro culture after
isolation using our gentle-enzymatic digestion method may allow for maturation of islets.