High-quality acinar cell isolation enables single-cell analysis of healthy and injured pancreas.

Acinar cells are the predominant cell type of the pancreas. However, in single-cell RNA sequencing datasets from healthy murine pancreas, acinar cells typically represent less than half of the total cells. Here, we developed a protocol that facilitates the rapid digestion of the pancreas into high-quality single cells from both healthy and caerulein-induced acute pancreatitis (AP) tissue. Under homeostatic conditions, acinar cells exhibited limited heterogeneity, with distinct subpopulations selectively and highly expressing protease genes or transcription factors, mirroring patterns observed in the healthy human pancreas. Moreover, 24 h after AP, acinar cells displayed heterogeneous expression of "ADM trypsinogens" and ductal markers. Additionally, this method enabled immune phenotyping via flow cytometry in a pancreatic ductal adenocarcinoma model. By overcoming the challenges associated with isolating acinar cells, our method offers an improvement over existing protocols for obtaining live single cells from the pancreas, which specifically caters to a comprehensive characterization of acinar cell populations.