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Modeling Primary Human Monocytes with the Trans–Differentiation Cell Line BLaER1

  • Moritz M. Gaidt
  • Francesca Rapino
  • Thomas Graf
  • Veit HornungEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1714)

Abstract

Monocytes and macrophages play a pivotal role in the induction and shaping of immune responses. Expressing a broad array of pattern recognition receptors (PRRs), monocytes and macrophages constitute an integral component of the innate branch of the immune system. Traditionally, the majority of innate immune sensing and signaling pathways have been studied in macrophages of the murine system. This is largely due to the fact that genetic loss-of-function studies are amenable in this species. On the other hand, human cell lines of the monocyte-macrophage cell lineage have been widely used to study myeloid cells in vitro. However, commonly utilized models (e.g., THP-1 cells) only mimic a limited spectrum of the immunobiology of primary human myeloid cells. Recently, we have explored the possibility to fill this gap with a human trans-differentiation cell culture system, in which lineage conversion from malignant B-lineage cells to monocytes/macrophages is caused by the inducible nuclear translocation of a C/EBPα transgene, BLaER1 cells. Using this model, we were able to characterize a novel inflammasome signaling entity that could not have been uncovered in the murine system or THP-1 cells. Here, we describe the handling of BLaER1 cells, providing a detailed protocol for their induced trans-differentiation. We also provide data to demonstrate the applicability of the BLaER1 monocyte/macrophage system to study phagocytosis and various PRR cascades in human cells.

Keywords

BLaER1 cell line BLaER1 monocytes Human monocytes Human macrophages Trans–differentiation Monocyte in vitro system PRR cascades Innate immunology 

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Moritz M. Gaidt
    • 1
  • Francesca Rapino
    • 2
    • 3
    • 4
  • Thomas Graf
    • 2
  • Veit Hornung
    • 1
    Email author
  1. 1.Gene Center and Department of BiochemistryLudwig-Maximilians-Universität MünchenMunichGermany
  2. 2.Center for Genomic RegulationUniversidad Pompeu Fabra and Institució Catalana de Recerca i Estudis AvançatsBarcelonaSpain
  3. 3.Department of Stem Cell and Regenerative BiologyHarvard UniversityCambridgeUSA
  4. 4.Harvard Stem Cell Institute, Harvard UniversityCambridgeUSA

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