Figure 1.

Sorting criteria of human B cell subsets from humanized NOD/SCID γc^null (hNSG) mice. (a) Single cell suspension was prepared from the bone marrow of a representative single hNSG mouse and cells in the lymphoid gate (not shown) were further gated for CD19⁺CD5⁻ expression (left). Cells were then plotted for surface IgM (sIgM) and CD10 expression (right), and single B cells from the early immature (sIgM⁻ CD10⁺) and immature (sIgM⁺CD10⁺) subsets were sorted. (b) PBMCs were prepared from 3 hNSG mice, all transplanted with the same donor and were gated for lymphoid population (not shown). These lymphocytes were gated on IgM⁺ cells (middle left) and further characterized as CD19⁺CD5⁻ or CD19⁺CD5⁺. Sorting was performed in the CD19⁺CD5⁻ gate to obtain single B cells from the new emigrant (CD10⁺CD27⁻) and mature (CD10⁺CD27⁻) B cell subsets (lower left). Similarly the CD5⁺CD19⁺ gated cells were single sorted to obtain CD5⁺ mature B cells (CD10⁺CD27⁻) (lower right). Single cell sortings were performed between 8–10 months post engraftment of the hNSG mice. All antibodies used were specific for human cell surface markers.