Optimizing Perfusion-Decellularization Methods of Porcine Livers for Clinical-Scale Whole-Organ Bioengineering

Figure 4

Recellularization of large-scale liver scaffolds prepared by three methods using rat primary hepatocytes after a 7-day perfusion culture. (a) H&E staining, (b) SEM micrographs of recellularized liver graft after 7 days in perfusion culture. (a) Scale bars: 100 μm; (b) arrows indicate hepatocytes, scale bars: 100 μm. (c) Albumin and (d) urea concentration in the culture medium of the collagen-Matrigel sandwich culture and in the perfusion culture medium of three groups of recellularized livers ( each group) during culture. Gene expression of (e) five liver-specific genes (Alb, Cyp1a1, Cyp1a2, HNF4α, and HNF6) and (f) six genes involved in the urea cycle (Nags, OTC, Cps1, Ass, Asl, and Arg1) were examined by qRT-PCR after 7-day culture. Collagen-sandwich group values served as calibrators to determine the relative expression of each target gene of each group. versus T-SDS group.

(a)

(b)

(c) Albumin synthesis

(d) Urea synthesis

(e)

(f)