Protocol to stress human primary pancreatic islets with diabetogenic factors for subsequent single-cell RNA sequencing

Type 1 diabetes (T1D) is an autoimmune disease that destroys insulin-secreting pancreatic β cells. Using human islets is instrumental to understanding the initiation and progression of T1D. Here, we present a protocol for treating cadaveric primary human islets with inflammatory, immune, and viral stressors. We detail Coxsackievirus B3 expansion, primary islet cell culture, and treatment with diabetogenic stressors, followed by procedures for single-cell RNA sequencing analysis. For complete execution details, please refer to Maestas et al. 1 and Veronese-Paniagua et al. 2