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Fig. 6 | Stem Cell Research & Therapy

Fig. 6

From: Donor antigen-primed regulatory T cells permit liver regeneration and phenotype correction in hemophilia A mouse by allogeneic bone marrow stem cells

Fig. 6

FVIII synthesis by donor-derived endothelial cells and Kupffer cells in HAT-AT liver. a Donor-derived endothelial cells indicated by white arrow (anti-GFP/donkey anti-mouse Alexa fluor 488 and anti-CD31/donkey anti-rat Alexa fluor 594) in HAT-AT liver (scale of 10 μm; 630 × 2.8 zoomed magnification). b Donor-derived Kupffer cells indicated by white arrow (anti-GFP/donkey anti-mouse Alexa fluor 488 and anti-F4/80/donkey anti-rat Alexa fluor 594) in HAT-AT liver (scale of 10 μm; 630 × 2.8 zoomed magnification). c Flow cytometric analysis of donor cells in non-parenchymal fraction of HAT-AT mouse liver after 1 month of transplantation. Representative dot-plots are given in the left panel, and quantitative analysis is shown in the right panel (n = 3). d FVIII(lc) gene expression in livers of HAT-AT, HA, and WT mice. e Immunocytochemical analysis of FVIII in endothelial and Kupffer cells from HA, WT, and HAT-AT mice (scale of 100 μm; 600× magnification). Relative cell fluorescence intensities are shown in the bar diagram on right. f Highly magnified confocal images (scale of 10 μm; 630× 2.8 zoomed magnification) show that donor-derived endothelial cells express FVIII indicated by white arrow (anti-GFP/donkey anti-mouse Alexafluor 488, anti-CD31/donkey anti-rat Alexafluor 594, anti-FVIII(lc)/donkey anti-rabbit Alexafluor 647). BMD bone marrow derived, DAPI 4′,6-diamidino-2-phenylindole, EC endothelial cell, GFP green fluorescent protein, FVIII factor VIII, FVIII(lc) factor VIII light chain, HA hemophilia A, HAT-AT hemophilia A mice transplanted with allogeneic and regulatory T cells, KC Kupffer cell, n number of mice in each experiment, NPC non-parenchymal cell, WT wild-type

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