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. 2023 Jan 20;14:1111627. doi: 10.3389/fendo.2023.1111627

Figure 1.

Figure 1

Surface expression of human and mouse CD45 on peripheral blood, bone marrow, splenocytes and bone marrow-derived osteoclasts of hu-BLT mice; comparison of immune cell percentages in human, WT mice and hu-BLT mice PBMCs. Hu-BLT mice acquired humanized hemato-lymphatic system by co-transplantation of human fetal liver and thymus fragments under the renal capsule of NOG/Scid γc-/- immune deficient NSG mice followed by the intravenous injection of autologous CD34+ hematopoietic cells. Eight weeks after tissue implantation, mice were euthanized and tissues were harvested to obtain single cell suspension. PBMCs, bone marrow (BM), and spleen of hu-BLT mice were analyzed for human and mice CD45+ immune cells using flow cytometric analysis (A). Hu-BLT mice BM isolated monocytes were used to generate osteoclasts (OCs) for 21 days as described in Materials and Methods section. On day 21, hu-BLT OCs were washed with 1 X PBS before the pictures were taken by Leica DMI 6000B inverted microscope. Hu-BLT OCs were then treated with Fast Garnet GBC and sodium nitrate (1:1), incubated at 37 degree C for one hour, cells were rinsed and treated with hematoxylin for two mins before pictures were taken by Leica DMI 6000B inverted microscope. Pre- and Post-TRAP-stained pictures are shown in the figure. Hu-BLT OCs were analyzed for human and mice CD45+ immune cells using flow cytometric analysis (B). Immune cell composition was determined in human PBMCs, hu-BLT PBMCs and B6 female WT mice PBMCs using flow cytometric analysis (C).