Figure 2 : Perfusable blood vessel formation and viable cardiac tissue fabrication.

From: In vitro fabrication of functional three-dimensional tissues with perfusable blood vessels

Figure 2

Blood vessel formation and tissue viability were evaluated among the four groups, with and without EC coculture, and with or without FGF-2 administration in the perfusion medium. Each group was indicated as EC (−) FGF (−), EC (−) FGF (+), EC (+) FGF (−) and EC (+) FGF (+). (ad) HE staining after a 3-day perfusion culture. Neither a luminal structure nor perfused black ink were detected in either the EC (−) FGF (−) group or in the EC (−) FGF (+) group. In the EC (+) FGF (−) group, a luminal structure was observed, but the black ink was hardly detected within it. In the EC (+) FGF (+) group, the infused black ink thoroughly reached the blood vessel structure within cell sheet layers (red arrows). (eh) CD31 staining for ECs. No blood vessel structure was observed within the cell sheet layers in either the EC (−) FGF (−) group or the EC (−) FGF (+) group. Significant tubular blood vessel structures (white arrows) were observed in the EC (+) FGF (−) group and in the EC (+) FGF (+) group. cell sheet (CS) indicates cell sheet constructs and vascular bed (VB) indicates vascular beds. (io) Luciferase-expressing triple-layer cell sheets were overlaid on the vascular beds. After continuous luciferin infusion, serial bioluminescent imaging of each group was analysed. The images captured at 45 min are shown (il). Representative dynamic tracing is demonstrated (m). The bioluminescent intensity at 45 and 90 min is shown (n,o). The intensity increased in the early phase in the EC (+) FGF (−) group and the EC (+) FGF (+) group. On the other hand, it increased in the late phase in the EC (−) FGF (−) group and the EC (−) FGF (+) group. FGF-2 accelerated the increasing rate, regardless of EC coculture. Data are shown as mean±s.d. (*P<0.05) (n=4). (p) Sarcomeric α-actinin staining (red) with nuclei staining (blue) revealed well-differentiated sarcomeres within the cell sheet construct in the EC (+) FGF (+) group. (q) HE staining showed that perfused black ink is occasionally detected EC(+) FGF(−) group.