Genetically modified human CD34+-derived microglial cells express marker or therapeutic genes in the brains of NOD/SCID mice. (A) Dot-plot analysis showing the percentage of human CD45+ cells expressing EGFP in mouse bone marrow. (B and C) EGFP-transduced UCB CD34+-derived cells in mouse brain. Nonramified (B) and ramified (C) cells express EGFP (in red, Cy3) and the human ANA marker (green, FITC) yielding a yellow staining in their nuclei (arrows). One ANA-positive cell (B, arrowhead) does not express EGFP. (D) Human UCB CD34+-derived cells differentiate into microglia expressing EGFP. Colocalization of EGFP (in red, Cy3) and Iba1 (in green, FITC) results in yellow staining (arrow). (E) CD68+ macrophages (in green, FITC, arrowheads) expressing recombinant ALD protein (punctate staining in red, Cy3, arrow) are present in the bone marrow of transplanted mice. (F) MPB CD34+ cells from ALD patient engrafted in mouse brain express recombinant ALD protein (punctate staining in red, Cy3, arrows) and the microglial RCA-1 lectin (in green, FITC). [Bars = 100 μm (B), 25 μm (C), and 10 μm (D-F).]