Efficient recellularisation of decellularised whole-liver grafts using biliary tree and foetal hepatocytes

Abstract

A whole-organ regeneration approach, using a decellularised xenogeneic liver as a scaffold for the construction of a transplantable liver was recently reported. Deriving suitable scaffolds was the first step towards clinical application; however, effective recellularisation remains to be achieved. This report presents a strategy for the improvement of the recellularisation process, using novel cell-seeding technique and cell source. We evaluated recellularised liver grafts repopulated through the portal vein or the biliary duct with mice adult hepatocytes or E14.5 foetal hepatocytes. More than 80% of the cells seeded through the biliary tree entered the parenchyma beyond the ductule-lining matrix barrier and distributed throughout the liver lobule. In contrast, about 20% of the cells seeded through the portal tree entered the parenchyma. The gene expression levels of foetal hepatocyte albumin, glucose 6-phosphatase, transferrin, cytokeratin 19, and gamma-glutamyl transpeptidase were increased in three-dimensional cultures in the native liver-derived scaffolds, and the activation of liver detoxification enzymes and formation of biliary duct-like structures were supported. The metabolic functions of liver grafts recellularised with different cell types were similar. These results suggest that biliary tree cell-seeding approach is promising, and that liver progenitor cells represent a good cell source candidate.

Vasculature reconstruction of decellularized liver scaffolds via gelatin‐based re‐endothelialization

“…Endothelial cells may enter the extravascular spaces through the altered basement membrane structures that were modified by the decellularization process as detergent‐based decellularization inevitably change/damage the ECM structures . In addition, endothelial cells that were injected to the decellularized liver scaffolds via extrahepatic bile ducts may also enter the extravascular spaces as previously shown . Shear stress has been shown to be an important modulator of endothelial cell morphology .…”

“…33,34 In addition, endothelial cells that were injected to the decellularized liver scaffolds via extrahepatic bile ducts may also enter the extravascular spaces as previously shown. 35 Shear stress has been shown to be an important modulator of endothelial cell morphology. 36,37 It is plausible that these cells may be subject to distinct shear stresses.…”

Liver tissue engineering: From implantable tissue to whole organ engineering

“…The fetal liver cells were seeded through the portal vein and vena cava but showed no accurate distribution of the various differentiated cells to the correct location within the liver lobules. Moreover, Ogiso et al60 demonstrated the existence of organ‐specific cell–ECM communication, which promotes the maturation of engrafted fetal hepatocytes into both hepatocyte and cholangiocyte lineages, without the addition of any prodifferentiation signals. In addition, it was found that using the biliary tree to seed the fetal hepatocytes resulted in a more accurate distribution of differentiated cells as well as an enhanced distribution of hepatocytes into the parenchyma compared to seeding through the vena cava.…”

Auxiliary xenotransplantation as an in vivo bioreactor—Development of a transplantable liver graft from a tiny partial liver

“…Numerous research projects using various agents to promote partial small liver grafts after LT have been implemented, all of which have only a limited effect for liver graft regeneration . Another approach is a challenging project to make transplantable livers using scaffolding and human embryonic stem cells (PSCs) through a tissue‐engineering technique . Because the structure is 3‐dimensional and several kinds of cells are involved intricately in the liver, projects attempting this approach may require more time.…”

“…[5][6][7] Another approach is a challenging project to make transplantable livers using scaffolding and human embryonic stem cells (PSCs) through a tissue-engineering technique. 8,9 Because the structure is 3-dimensional and several kinds of cells are involved intricately in the liver, projects attempting this approach may require more time.…”