Functional behavior of ex vivo–generated TPs in NOD/SCID mice. (A) Analyses of human erythroid cells in peripheral blood of NOD/SCID mice receiving the TP. The mean ± SD of the percentage of human erythroid cells in the blood of NOD/SCID mice on serial days as described by double staining with LDS (nuclei) and human GPA. LDS⁺GPA⁺ represents human EPCs (dotted line) and LDS⁻GPA⁺ represents erythrocytes (solid line). Evidence of human EPCs and/or erythrocytes was not observed in control mice (black line indicates LDS⁻GPA⁺; and LDS⁺GPA⁺, gray line). Two to 3 mice were analyzed in each group (n = 2). (B) Representative histogram of human erythroid cells present on day 5 in the peripheral blood of NOD/SCID mouse. Similar results were obtained in an additional experiment. (C) Detection of human globins (β and γ) in the NOD/SCID mouse blood 15 days following the infusion of the transfusion product. Primers of mouse α-globin, human β and γ globins were mixed together and expression of globin mRNA was analyzed from cDNA prepared from mouse blood on day 15 by RT-PCR. Hu-β (212 bp), hu-γ (165 bp), RhD antigen (90 bp), and mouse-α (122 bp) gene products are shown on a 1.8% agarose gel. Lanes M1 and M2 contain DNA marker (a 100-bp ladder and 50-bp ladder); lane 1, blood from control NOD/SCID mouse; and lane 2, blood from NOD/SCID mouse infused with the transfusion product. One representative of 2 experiments is shown. (D) Human cell engraftment in the marrow of NOD/SCID mice. (i) Mouse bone marrow was harvested and analyzed on day 15 (NOD/SCID mice, n = 2) and (ii) in separate series of experiment NOD/SCIDγc^null mice day 30 (n = 3) after the infusion of the TP.