American Society of Hematology

Figure 1

$Figure 1. E11.5 AGM-derived repopulating cells do not express CD150 or CD48. (A) E11.5 embryos before (i) and after (ii) AGM dissection. The image was captured at room temperature by the use of a Leica MZ16 scope and a Nikon COOLPIX camera. Total magnification ×400. Arrows denote the aorta. (B) Flow cytometry analysis of CD48, CD150, and CD41 expression on AGM cells. (C) To assess the in vivo hematopoietic repopulating potential of cells isolated from E11.5 AGM, E12.5 placenta, E14.5 FL, and WBM, donor cells from each tissue were isolated from CD45.2 C57BL/6 mice, fractionated via FACS, and then transplanted into irradiated CD45.1 C57BL/6 mice along with 2.5 × 105 competitor WBM cells isolated from CD45.1/CD45.2 C57BL/6 mice. Recipient PB mice were then analyzed every 4 weeks for the presence of CD45.2+ (ie, donor-derived) cells. (D) A representative mouse 7 months after transplant with CD150−CD48− E11.5 AGM-derived (CD45.2+) B cells (IgM+/CD19+), T cells (CD3+ and/or CD4+/CD8+), and myeloid cells (Gr-1+ and/or Mac-1+).$

E11.5 AGM-derived repopulating cells do not express CD150 or CD48. (A) E11.5 embryos before (i) and after (ii) AGM dissection. The image was captured at room temperature by the use of a Leica MZ16 scope and a Nikon COOLPIX camera. Total magnification ×400. Arrows denote the aorta. (B) Flow cytometry analysis of CD48, CD150, and CD41 expression on AGM cells. (C) To assess the in vivo hematopoietic repopulating potential of cells isolated from E11.5 AGM, E12.5 placenta, E14.5 FL, and WBM, donor cells from each tissue were isolated from CD45.2 C57BL/6 mice, fractionated via FACS, and then transplanted into irradiated CD45.1 C57BL/6 mice along with 2.5 × 10⁵ competitor WBM cells isolated from CD45.1/CD45.2 C57BL/6 mice. Recipient PB mice were then analyzed every 4 weeks for the presence of CD45.2⁺ (ie, donor-derived) cells. (D) A representative mouse 7 months after transplant with CD150⁻CD48⁻ E11.5 AGM-derived (CD45.2⁺) B cells (IgM⁺/CD19⁺), T cells (CD3⁺ and/or CD4⁺/CD8⁺), and myeloid cells (Gr-1⁺ and/or Mac-1⁺).

This Feature Is Available To Subscribers Only