Myeloid Engraftment in Humanized Mice: Impact of Granulocyte-Colony Stimulating Factor Treatment and Transgenic Mouse Strain

Abstract

Poor myeloid engraftment remains a barrier to experimental use of humanized mice. Focusing primarily on peripheral blood cells, we compared the engraftment profile of NOD-scid-IL2Rγc(-/-) (NSG) mice with that of NSG mice transgenic for human membrane stem cell factor (hu-mSCF mice), NSG mice transgenic for human interleukin (IL)-3, granulocyte-macrophage-colony stimulating factor (GM-CSF), and stem cell factor (SGM3 mice). hu-mSCF and SGM3 mice showed enhanced engraftment of human leukocytes compared to NSG mice, and this was reflected in the number of human neutrophils and monocytes present in these strains. Importantly, discrete classical, intermediate, and nonclassical monocyte populations were identifiable in the blood of NSG and hu-mSCF mice, while the nonclassical population was absent in the blood of SGM3 mice. Granulocyte-colony stimulating factor (GCSF) treatment increased the number of blood monocytes in NSG and hu-mSCF mice, and neutrophils in NSG and SGM3 mice; however, this effect appeared to be at least partially dependent on the stem cell donor used to engraft the mice. Furthermore, GCSF treatment resulted in a preferential expansion of nonclassical monocytes in both NSG and hu-mSCF mice. Human tubulointerstitial CD11c(+) cells were present in the kidneys of hu-mSCF mice, while monocytes and neutrophils were identified in the liver of all strains. Bone marrow-derived macrophages prepared from NSG mice were most effective at phagocytosing polystyrene beads. In conclusion, hu-mSCF mice provide the best environment for the generation of human myeloid cells, with GCSF treatment further enhancing peripheral blood human monocyte cell numbers in this strain.

Functional compensation between hematopoietic stem cell clones in vivo

“…We found significant activation of genes involved in diseases, biological functions, and networks that are related to lymphopoietic compensation ( Fig EV3, and Tables EV1 and EV2). In the NSG co-transplantation group, we found an enrichment for genes involved in natural killer (NK) cell proliferation, including Il7r [27], Slamf6 [29], and Axl [30] (Fig 5B), consistent with the previous findings that HSCs derived from NSG mice are unable to produce NK cells [31,32]. These data suggest that compensation for lymphopoietic deficiency may be communicated and determined at the HSC level.…”

Identification of an Early Unipotent Neutrophil Progenitor with Pro-tumoral Activity in Mouse and Human Bone Marrow

“…The triple transgenic NSG-M3 mice are immunodeficient NOD scid gamma (NSG) mice that express the human cytokines Interleukin 3 (IL-3), granulocyte/macrophage-stimulating factor (GM-CSF) and SCF, also known as KITLG. This mouse model supports stable engraftment of the human hematopoietic system, including the myeloid lineage (Billerbeck et al, 2011; Coughlan et al, 2016). The two subsets were isolated from fresh human bone marrow by FACS using the sorting panel in Figure 5 and transferred into two groups of recipient NSG-M3 mice.…”

Comparison of Human Hematopoietic Reconstitution in Different Strains of Immunodeficient Mice

“…Transgenes coding for human cytokines are responsible for the elevated levels of myelopoiesis observed in NSG-3GS and hSCF-NSG mice [8,19]. Conversely, the high frequency of B-lymphopoiesis in NSG mice was due to weaker myelopoiesis in these mice.…”