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Fig. 4. CEACAM1 expression in new endothelial sprouts. Immunostaining for
CEACAM1 on sections of embedded HITA rings demonstrate the presence of
CEACAM1-positive cells within the lumen (A) and in the outside of HITA
rings (B). Mature endothelial cells lining HITA lumen and not involved
in the sprouting remain negative for CEACAM1 (arrows). CD34 immunostaining of
a serial section following the section in B confirms the involvement of
CD34+ cells (arrows) in formation of these new capillaries at the
border between vascular wall and the collagen gel (C). CEACAM1 is not
present in the quiescent endothelial cells lining HITA lumen or in the
vasculogenic zone of HITA when ring assay was not performed (D). (E-H)
GFP marking of HITA-wall cells in ring assay. The injection of AdV5-GFP into a
zone of the outer layer of the HITA demonstrates GFP-marked cells within and
migrating out from the vascular wall (E). Higher magnification shows
the sprouts from the vascular wall into the collagen gel (F). Focusing
on another level demonstrates the formation of vascular like channel (arrows)
at the border between the vascular wall and collagen gel corresponding to the
vascular channels presented in B and C (G). No such sprouting is
visible in the control ring without GFP-AdV5 (H).
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