Figure 1.
Figure 1. Engraftment of ex vivo expanded FL hematopoietic cells in NOD/SCID mice. (A) Summary of the levels of human cell engraftment in the BM of NOD/SCID mice given transplants with various doses of FL no. 841 cells, unexpanded (upper panel) or expanded for 28 days (lower panel). The percentage of human cells in the individual recipients was determined 8 weeks after transplantation by flow cytometry analysis of BM cells expressing human CD45. The dotted lines separate engrafted from nonengrafted animals, as confirmed by Southern blot analysis. More detailed analyses of the engraftment of expanded cells from this FL specimen are shown below. (B) Flow cytometry analysis of a highly engrafted representative mouse given transplants with 5 × 106 expanded FL no. 841 cells. About 56% of the cells in this mouse BM stained positively for the human pan-leukocyte marker CD45 (upper left panel, gate R1, and upper right panel, region M1); these human cells, once backgated to a forward/side scatter (FSC/SSC) plot, mostly fell within a typical viable (but heterogeneous) cell population (lower left panel). The middle and bottom right panels show a CD45 staining of a control (not transplanted) NOD/SCID mouse BM, and an isotypic control (mouse IgG1) of a positively engrafted NOD/SCID mouse, respectively. Both panels show only a few rare events falling in gate R1. (C) Representative examples of positive engraftment of NOD/SCID mice given transplants with 1 to 10 × 106 expanded FL no. 841 cells as indicated. Human cells appear in gate R1, and their percentages (of total BM cells) are shown in the respective panels. (D) Southern blot analysis of human cell engraftment in the BM of individual NOD/SCID mice given transplants with the indicated doses (0.25 to 10 × 106 TNCs) of FL no. 841 cells following 28 days of expansion. EcoRI-digested DNA (4 μg) was loaded in each lane, and the blot was hybridized to a human chromosome 17-specific α-satellite probe as described.9 Positive engraftment is indicated by plus signs above the respective lanes. Human/mouse (H/M) DNA controls are given as percentage of human DNA. M lanes were loaded with DNA molecular weight markers.

Engraftment of ex vivo expanded FL hematopoietic cells in NOD/SCID mice. (A) Summary of the levels of human cell engraftment in the BM of NOD/SCID mice given transplants with various doses of FL no. 841 cells, unexpanded (upper panel) or expanded for 28 days (lower panel). The percentage of human cells in the individual recipients was determined 8 weeks after transplantation by flow cytometry analysis of BM cells expressing human CD45. The dotted lines separate engrafted from nonengrafted animals, as confirmed by Southern blot analysis. More detailed analyses of the engraftment of expanded cells from this FL specimen are shown below. (B) Flow cytometry analysis of a highly engrafted representative mouse given transplants with 5 × 106 expanded FL no. 841 cells. About 56% of the cells in this mouse BM stained positively for the human pan-leukocyte marker CD45 (upper left panel, gate R1, and upper right panel, region M1); these human cells, once backgated to a forward/side scatter (FSC/SSC) plot, mostly fell within a typical viable (but heterogeneous) cell population (lower left panel). The middle and bottom right panels show a CD45 staining of a control (not transplanted) NOD/SCID mouse BM, and an isotypic control (mouse IgG1) of a positively engrafted NOD/SCID mouse, respectively. Both panels show only a few rare events falling in gate R1. (C) Representative examples of positive engraftment of NOD/SCID mice given transplants with 1 to 10 × 106 expanded FL no. 841 cells as indicated. Human cells appear in gate R1, and their percentages (of total BM cells) are shown in the respective panels. (D) Southern blot analysis of human cell engraftment in the BM of individual NOD/SCID mice given transplants with the indicated doses (0.25 to 10 × 106 TNCs) of FL no. 841 cells following 28 days of expansion. EcoRI-digested DNA (4 μg) was loaded in each lane, and the blot was hybridized to a human chromosome 17-specific α-satellite probe as described.9  Positive engraftment is indicated by plus signs above the respective lanes. Human/mouse (H/M) DNA controls are given as percentage of human DNA. M lanes were loaded with DNA molecular weight markers.

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