Protocol to study regeneration in human pancreatic slices using viral tracing, extended culture, dissociation, and functional Ca2+ influx techniques

Human pancreatic slices provide an organotypic platform that preserves native cell interactions for functional studies under near-physiological conditions. We present a protocol to regenerating human pancreatic slices, including lentiviral and adenoviral transduction for lineage tracing and a dual-reporter HIP-Cre system to label neogenic β-cells. The workflow includes tissue dissociation for single-cell analysis and calcium influx measurements to assess glucose responsiveness, enabling identification, tracking, and functional evaluation of newly formed β-cells. For complete details on the use and execution of this protocol, please refer to Doke et al.¹.