An optimized protocol for isolation of murine pancreatic single cells with high yield and purity

• Published in: STAR protocols Vol. 5; no. 1; p. 102836
• Main Authors: Wu, Feijing, Jiang, Zhengyu, Qian, Jin, Kobayashi, Hiroki, Waterbury, Quin T., White, Ruth A., Ochiai, Yosuke, Zhi, Xiaofei, Tu, Ruhong, Zheng, Biyun, Shi, Qiongyu, Zamechek, Leah B., Wang, Timothy C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.03.2024; Elsevier
• Subjects: Cell Biology; Cell isolation; Flow Cytometry; Organoids; Protocol; Single Cell; Single Cell; Flow Cytometry; Organoids; Cell isolation; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.102836

Here, we present a protocol for rapidly isolating single cells from the mouse pancreas, minimizing damage caused by digestive enzymes in exocrine cells. We guide you through steps to optimize the dissection sequence, enzyme composition, and operational procedures, resulting in high yields of viable pancreatic single cells. This protocol can be applied across a wide range of research areas, including single-cell sequencing, gene expression profiling, primary cell culture, and even the development of spheroids or organoids. For complete details on the use and execution of this protocol, please refer to Jiang et al. (2023).1 [Display omitted] •Isolate pancreatic tissue while minimizing enzyme damage•Conduct an efficient pancreatic tissue digestion procedure•Label and isolate specific cell types using flow cytometry•Culture and analyze isolated pancreatic single cells Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for rapidly isolating single cells from the mouse pancreas, minimizing damage caused by digestive enzymes in exocrine cells. We guide you through steps to optimize the dissection sequence, enzyme composition, and operational procedures, resulting in high yields of viable pancreatic single cells. This protocol can be applied across a wide range of research areas, including single-cell sequencing, gene expression profiling, primary cell culture, and even the development of spheroids or organoids.