Generation of Humanized Mice for Analysis of Human Dendritic Cells

  • Yasuyuki SaitoEmail author
  • Jana M. Ellegast
  • Markus G. Manz
Part of the Methods in Molecular Biology book series (MIMB, volume 1423)


Transplantation of human CD34+ hematopoietic stem and progenitor cells into severe immunocompromised newborn mice allows the development of a human hemato-lymphoid system (HHLS) including dendritic cells (DCs) in vivo. Therefore, it can be a powerful tool to study human DC subsets, residing in different lymphoid and nonlymphoid organs. We have recently generated novel mouse strains called human cytokine knock-in mice in which human versions of several cytokines are knocked into Rag2−/−γC−/− strains. In addition, human SIRPα, which is a critical factor to prevent donor cell to be eliminated by host macrophages, is expressed as transgene. These mice efficiently support human myeloid cell development and, indeed, allow the analysis of three major subsets of human DC lineages, plasmacytoid DCs and CD1c+ and CD141+ classical DCs. Moreover, these strains also support cytokine-mobilized peripheral blood CD34+ cell engraftment and subsequent DC development. Here we describe our standard methods to characterize DCs developed in human cytokine knock-in mice.

Key words

Humanized mouse CD34+ hematopoietic stem and progenitor cell Stem cell transplantation Human dendritic cells 


  1. 1.
    Shortman K, Liu Y-J (2002) Mouse and human dendritic cell subtypes. Nat Rev Immunol 2:151–161CrossRefPubMedGoogle Scholar
  2. 2.
    Palucka K, Banchereau J (2013) Human dendritic cell subsets in vaccination. Curr Opin Immunol 25:396–402CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Merad M, Sathe P, Helft J et al (2013) The dendritic cell lineage: ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting. Ann Rev Immunol 31:563–604CrossRefGoogle Scholar
  4. 4.
    Ishikawa F, Niiro H, Iino T et al (2007) The developmental program of human dendritic cells is operated independently of conventional myeloid and lymphoid pathways. Blood 110:3591–3660CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Tanaka S, Saito Y, Kunisawa J et al (2012) Development of mature and functional human myeloid subsets in hematopoietic stem cell-engrafted NOD/SCID/IL2rγKO Mice. J Immunol 188:6145–6155CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Poulin LF, Reyal Y, Uronen-Hansson H et al (2012) DNGR-1 is a specific and universal marker of mouse and human Batf3-dependent dendritic cells in lymphoid and nonlymphoid tissues. Blood 119:6052–6062CrossRefPubMedGoogle Scholar
  7. 7.
    Rongvaux A, Willinger T, Martinek J et al (2014) Development and function of human innate immune cells in a humanized mouse model. Nat Biotech 32:364–372CrossRefGoogle Scholar
  8. 8.
    Takenaka K, Prasolava TK, Wang JCY et al (2007) Polymorphism in Sirpa modulates engraftment of human hematopoietic stem cells. Nat Immunol 8:1313–1323CrossRefPubMedGoogle Scholar
  9. 9.
    Strowig T, Rongvaux A, Rathinam C et al (2011) Transgenic expression of human signal regulatory protein alpha in Rag2-/- gamma c-/- mice improves engraftment of human hematopoietic cells in humanized mice. Proc Natl Acad Sci U S A 108:13218–13223CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Ziegler P, Manz MG (2007) Mouse models for human hemato-lymphopoiesis. Curr Protocol Toxicol. Chapter 2, Unit2.13Google Scholar
  11. 11.
    van Lent AU, Centlivre M, Nagasawa M et al (2010) In vivo modulation of gene expression by lentiviral transduction in “human immune system” Rag2-/- gamma c -/- mice. Methods Mol Biol 595:87–115CrossRefPubMedGoogle Scholar
  12. 12.
    Gimeno R, Weijer K, Voordouw A et al (2004) Monitoring the effect of gene silencing by RNA interference in human CD34+ cells injected into newborn RAG2-/- gammac-/- mice: functional inactivation of p53 in developing T cells. Blood 104:3886–3893CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Yasuyuki Saito
    • 1
    • 2
    Email author
  • Jana M. Ellegast
    • 1
  • Markus G. Manz
    • 1
  1. 1.Division of HematologyUniversity Hospital ZurichZurichSwitzerland
  2. 2.Division of Molecular and Cellular Signaling, Department of Biochemistry and Molecular BiologyKobe University Graduate School of MedicineKobeJapan

Personalised recommendations