Protocol for the induction of innate immune memory in human smooth muscle cells and endothelial cells in vitro

STAR Protoc. 2024 Mar 15;5(1):102776. doi: 10.1016/j.xpro.2023.102776. Epub 2023 Dec 19.

Abstract

Non-immune cells, like innate immune cells, can develop a memory-like phenotype in response to priming with microbial compounds or certain metabolites, which enables an enhanced response to a secondary unspecific stimulus. This paper describes a step-by-step protocol for the induction and analysis of trained immunity in human endothelial and smooth muscle cells. We then describe steps for cell culture with cryopreserved vascular cells, subcultivation, and induction of trained immunity. We then provide detailed procedures for downstream analysis using ELISA and qPCR. For complete details on the use and execution of this protocol, please refer to Sohrabi et al. (2020)1 and Shcnack et al.2.

Keywords: Cell Biology; Immunology; Molecular Biology.

MeSH terms

  • Cell Culture Techniques
  • Endothelial Cells*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Myocytes, Smooth Muscle
  • Trained Immunity*