Isolation and phenotypic characterization of human and nonhuman primate intestinal lamina propria mononuclear cells

• Published in: STAR protocols 2022-12, Vol.3 (4), p.101815-101815, Article 101815
• Main Authors: Benmeziane, Keltouma, Delache, Benoit, Langlois, Sébastien, Scarlatti, Gabriella, Le Grand, Roger, Cavarelli, Mariangela
• Format: Article
• Language: English
• Subjects: Animals; Cell isolation; Flow Cytometry - methods; Flow Cytometry/Mass Cytometry; Humans; Immunology; Intestinal Mucosa; Leukocytes; Life Sciences; Primates; Protocol
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101815

Isolation of viable immune cells from tissues is critically important to characterize cellular and molecular processes during homeostasis and disease. Here, we provide an optimized protocol to achieve high yields of viable intestinal lamina propria mononuclear cells (LPMCs). We describe steps for intestinal tissue collection from humans and nonhuman primates, followed by mechanical disruption and enzymatic digestion. Furthermore, we detail characterization of the mononuclear phagocyte (MP) subtypes by flow cytometry analysis. The protocol is repeatable and scalable for downstream applications. For complete details on the use and execution of this protocol, please refer to Cavarelli et al. (2022). [Display omitted] •Collection and identification of the portions of the lower intestinal tract•Detailed protocol for intestinal lamina propria mononuclear cells isolation•Detailed protocol for lamina propria mononuclear cells staining•Procedures to identify mononuclear phagocyte subsets by flow cytometry Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Isolation of viable immune cells from tissues is critically important to characterize cellular and molecular processes during homeostasis and disease. Here, we provide an optimized protocol to achieve high yields of viable intestinal lamina propria mononuclear cells (LPMCs). We describe steps for intestinal tissue collection from humans and nonhuman primates, followed by mechanical disruption and enzymatic digestion. Furthermore, we detail characterization of the mononuclear phagocyte (MP) subtypes by flow cytometry analysis. The protocol is repeatable and scalable for downstream applications.