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

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


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.


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


  1. 1.
    Mak TW, Penninger JM, Ohashi PS (2001) Knockout mice: a paradigm shift in modern immunology. Nat Rev Immunol 1:11–19. CrossRefPubMedGoogle Scholar
  2. 2.
    Khare S, Dorfleutner A, Bryan NB et al (2012) An NLRP7-containing inflammasome mediates recognition of microbial lipopeptides in human macrophages. Immunity 36:464–476. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Mestas J, Hughes CCW (2004) Of mice and not men: differences between mouse and human immunology. J Immunol 172:2731–2738. CrossRefPubMedGoogle Scholar
  4. 4.
    Cavlar T, Deimling T, Ablasser A et al (2013) Species-specific detection of the antiviral small-molecule compound CMA by STING. EMBO J 32:1440–1450. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Schroder K, Irvine KM, Taylor MS et al (2012) Conservation and divergence in toll-like receptor 4-regulated gene expression in primary human versus mouse macrophages. Proc Natl Acad Sci 109:E944–E953. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Tsuchiya S, Yamabe M, Yamaguchi Y et al (1980) Establishment and characterization of a human acute monocytic leukemia cell line (THP-1). Int J Cancer 26:171–176. CrossRefPubMedGoogle Scholar
  7. 7.
    Sundström C, Nilsson K (1976) Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer 17:565–577. CrossRefPubMedGoogle Scholar
  8. 8.
    Gaidt MM, Ebert TS, Chauhan D et al (2016) Human monocytes engage an alternative inflammasome pathway. Immunity 44:833–846. CrossRefPubMedGoogle Scholar
  9. 9.
    Chanput W, Peters V, Wichers H (2015) THP-1 and U937 cells. In: Verhoeckx K, Cotter P, López-Expósito I et al (eds) Impact food bioact. Heal. Vitr. Ex vivo model. Springer International Publishing, Cham, pp 147–159Google Scholar
  10. 10.
    Chanput W, Mes J, Vreeburg RA et al (2010) Transcription profiles of LPS-stimulated THP-1 monocytes and macrophages: a tool to study inflammation modulating effects of food-derived compounds. Food Funct 1:254–261. CrossRefPubMedGoogle Scholar
  11. 11.
    Klinken SP, Alexander WS, Adams JM (1988) Hemopoietic lineage switch: v-raf oncogene converts Emu-myc transgenic B cells into macrophages. Cell 53:857–867. CrossRefPubMedGoogle Scholar
  12. 12.
    Borzillo GV, Ashmun R, Sherr CJ (1990) Macrophage lineage switching of murine early pre-B lymphoid cells expressing transduced fms genes. Mol Cell Biol 10:2703–2714. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Xie H, Ye M, Feng R, Graf T (2004) Stepwise reprogramming of B cells into macrophages. Cell 117:663–676. CrossRefPubMedGoogle Scholar
  14. 14.
    Bussmann LH, Schubert A, Vu Manh TP et al (2009) A robust and highly efficient immune cell reprogramming system. Cell Stem Cell 5:554–566. CrossRefPubMedGoogle Scholar
  15. 15.
    Rapino F, Robles EF, Richter-Larrea JA et al (2017) C/EBPa induces highly efficient macrophage transdifferentiation of B lymphoma and leukemia cell lines and impairs their tumorigenicity. Cell Rep 19(6):1981.
  16. 16.
    Schlee M, Roth A, Hornung V et al (2009) Recognition of 5′ triphosphate by RIG-I helicase requires short blunt double-stranded RNA as contained in panhandle of negative-strand virus. Immunity 31:25–34. CrossRefPubMedPubMedCentralGoogle Scholar

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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