STAR Protocols (Jun 2022)

Isolation of human intrahepatic leukocytes for phenotypic and functional characterization by flow cytometry

  • Stephanie Kucykowicz,
  • Oliver E. Amin,
  • Alice R. Burton,
  • Leo Swadling,
  • Nathalie M. Schmidt,
  • Nekisa Zakeri,
  • Jessica Davies,
  • Gloryanne Aidoo-Micah,
  • Kerstin A. Stegmann,
  • Nicholas J. Easom,
  • Anna Jeffery-Smith,
  • Mala K. Maini,
  • Laura J. Pallett

Journal volume & issue
Vol. 3, no. 2
p. 101356

Abstract

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Summary: With the growing appreciation of tissue-resident immunity, studying tissue-specific immune cells contributing to both homeostasis and disease is imperative. Here, we provide a protocol for the isolation of human intrahepatic leukocytes (IHL) maximizing viability, purity, and yield. Our protocol is scalable by tissue weight, allowing for reproducible and efficient IHL liberation suitable for functional characterization, cell isolation, and profiling by flow (or mass) cytometry. Furthermore, we provide a “guide” to determine an expected IHL yield per gram of tissue processed.For complete details on the use and execution of this protocol, please refer to Stegmann et al. (2016), Pallett et al. (2017), Easom et al. (2018), Swadling et al. (2020), Pallett et al. (2020), and Zakeri et al. (2022).

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