Diabetes represents a major endemic disease throughout the world, and different therapeutic methods are used to treat the disease. Xenotransplantation of pig islet cells is a potential treatment for type 1 diabetes, but studies of protein expression in distinct islet cells are rare. ZnT8, a member of the slc30A gene family, is involved in islet endocrine hormone release and is a diabetes auto-antigen, raising the question of whether ZnT8 expression is regulated similarly in pig and human pancreas. We used nested RT-PCR to detect ZnT8 expression in pig pancreas and polyclonal antibody to examine possible co-localization with other islet hormones. Immunohistochemistry of sequential serial sections as well as double immunostaining of pancreatic tissues with antibodies against ZnT8, insulin, glucagon, and somatostatin shows that pig ZnT8 is exclusively co-expressed in insulin-producing, but not in glucagon- or somatostatin-producing cells. The absence of ZnT8 in glucagon-producing cells in pig islets indicates that zinc homeostasis is mediated by a different cellular mechanism compared with human islet cells. Our findings provide important information about the cell-type-specific expression of ZnT8 in porcine islet cells, which should be taken into account when evaluating different xenotransplantation approaches.