American Society of Hematology

Figure 5

$Figure 5. WAT anastomosis with syngeneic EC networks. (A) The perfused fraction of engrafted vessels was assessed at days 3, 6, 9, and 13 after implantation. PMBECs anastomosed with the host vasculature much more rapidly and efficiently than HUVECs. Three to 5 images (1280 μm × 1280 μm) from each of 3 animals were analyzed for each group at each time point. (B) Dynamics at the interface of engrafted and host networks. PMBECs from Tie2-GFP/Rag1−/− mice (green) migrate to and wrap around host vessels (red, mCD31-X647 injected intravenously) of a Rag1−/− mouse. Note that because perfused host vessels, perfused PMBEC vessels, and wrapping PMBECs are all labeled with the injected mCD31-X647 antibody, the replacement of host vessels by implanted PMBECs can only be identified when a vessel that was previously red (ie, unwrapped and perfused host vessel) has become yellow (PMBECs with or without regressing host vessel segments inside). The illustration in the right-hand column shows the major segments involved and indicates the locations of attachment to nearby networks consisting only of implanted PMBECs (green arrows) or host ECs (red arrows). This column also shows the fate of various segments in the tracked host vascular network. Once the orange section has been wrapped by the engrafted PMBECs, the adjacent purple segment regresses and the blue segment dilates. By day 11, there is extensive fluid communication between the implanted and host networks because of WAT anastomosis in this region. Intravital confocal images; 20×/0.4 NA air objective at 2.0× digital zoom; scale bar represents 50 μm. (C) Ten-day-old implant showing remnants of host vessels from the Tie2-GFP/Rag1−/− mouse (green, filled arrowheads) within vessels formed from implanted PMBECs collected from Rag1−/− mice (red, mCD31 staining). Note that mCD31 stains both the host and the implanted ECs. “Box” shows the area within the white box, but with fewer z-slices included in the projection to eliminate some of the overlying and underlying vessels. Whole-mount IHC stain; 60×/1.1 NA oil objective; scale bar represents 30 μm.$

WAT anastomosis with syngeneic EC networks. (A) The perfused fraction of engrafted vessels was assessed at days 3, 6, 9, and 13 after implantation. PMBECs anastomosed with the host vasculature much more rapidly and efficiently than HUVECs. Three to 5 images (1280 μm × 1280 μm) from each of 3 animals were analyzed for each group at each time point. (B) Dynamics at the interface of engrafted and host networks. PMBECs from Tie2-GFP/Rag1^−/− mice (green) migrate to and wrap around host vessels (red, mCD31-X647 injected intravenously) of a Rag1^−/− mouse. Note that because perfused host vessels, perfused PMBEC vessels, and wrapping PMBECs are all labeled with the injected mCD31-X647 antibody, the replacement of host vessels by implanted PMBECs can only be identified when a vessel that was previously red (ie, unwrapped and perfused host vessel) has become yellow (PMBECs with or without regressing host vessel segments inside). The illustration in the right-hand column shows the major segments involved and indicates the locations of attachment to nearby networks consisting only of implanted PMBECs (green arrows) or host ECs (red arrows). This column also shows the fate of various segments in the tracked host vascular network. Once the orange section has been wrapped by the engrafted PMBECs, the adjacent purple segment regresses and the blue segment dilates. By day 11, there is extensive fluid communication between the implanted and host networks because of WAT anastomosis in this region. Intravital confocal images; 20×/0.4 NA air objective at 2.0× digital zoom; scale bar represents 50 μm. (C) Ten-day-old implant showing remnants of host vessels from the Tie2-GFP/Rag1^−/− mouse (green, filled arrowheads) within vessels formed from implanted PMBECs collected from Rag1^−/− mice (red, mCD31 staining). Note that mCD31 stains both the host and the implanted ECs. “Box” shows the area within the white box, but with fewer z-slices included in the projection to eliminate some of the overlying and underlying vessels. Whole-mount IHC stain; 60×/1.1 NA oil objective; scale bar represents 30 μm.

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