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Systemic multilineage engraftment in mice after in utero transplantation with human hematopoietic stem cells

Russell G. Witt, Emily M. Kreger, Laura B. Buckman, Patriss W. Moradi, Phong T. Ho, S. Christopher Derderian, Perry Tsai, Chris Baker, Nathaniel Schramm, Rachel Cleary, J. Victor Garcia and Tippi C. MacKenzie

Data supplements

Article Figures & Data

Figures

  • Figure 1.

    Stable, multilineage chimerism in humanized mice after in utero transplantation of human cord blood CD34+cells. (A) Experimental design for in utero transplantation timing and measurements of blood chimerism levels. Arrows indicate time points for peripheral blood chimerism checks. (B) Representative gating strategy of peripheral blood to detect human CD45+ cells. (C) Percentage of human CD45+ cells in peripheral blood over time (n = 25, *P < .05, **P < .01 by Student t test comparing chimerism levels each week to the initial 4-week level). (D) Representative gating strategy for lineage-specific chimerism, FoxP3+ cells, and confirmation of the presence of CD34+ cells within the bone marrow. Chimeric mice demonstrated different relative proportions of granulocytes, monocytes, B cells, and T cells. (E) Compiled lineage data in 10 individual mice with evidence of CD34+ cells in the bone marrow (n = 10). CB, cord blood; SSC-A, side scatter.

  • Figure 2.

    Human CD45+cells show systemic multilineage engraftment with increased engraftment rates following ACK2 treatment. (A) Percentage of human CD45+ cells in lymphoid tissues of chimeric mice and peripheral blood. *P < .05, ***P < .001, ****P < .0001 (analysis of variance with Tukey’s honest significance test). (B) Immunohistochemistry of harvested spleen from a T-cell–dominant mouse demonstrating organized lymphoid follicles. (original magnification ×10; immunohistochemical analysis was performed on fixed/paraffin-embedded tissue sections for human CD45+, CD3+, CD4+, CD8+, CD20+, and CD68+ then counterstained with hematoxylin) (C) Effect of ACK2 treatment on the rates of engraftment (the proportion of surviving animals with ≥1% chimerism in any tissue). (D) Overall chimerism levels. (E-H) NSG mice treated with in utero transplantation of human CD34+ cells demonstrate populations of human cells, within the lung alveolar and bronchiole tissue (E), small and large bowel (F), female reproductive tract (G), and brain (H) (original magnification ×40; immunohistochemical analysis was performed on fixed/paraffin-embedded tissue sections for human CD45+, CD3+, CD4+, CD8+, CD68+, and CD20+ and counterstained with hematoxylin). BM, bone marrow; LN, lymph nodes; PB, peripheral blood.