Bioartificial liver - decellularization and the subsequent recellularization of the rat liver

Abstract

Introduction: Bioartificial livers (B.A.L) could become a possible future alternative to the liver transplant. B.A.L would be used as a temporary transplant (to sustain a critically ill patient until a suitable donor organ becomes available) or a permanent transplant. It takes the form of closed, ex vivo systems containing functional liver cells grown on a synthetic matrix. The aim of the present study is to obtain a bioartificial rat liver by consequently using of the decellularization and recellularization specific methods. Materials and methods: 20 rats ranging in age from 6 weeks to 3 months have been used for obtaining intact rat liver. The obtained liver was mounted on a decellularization apparatus for perfusion. The portal vein was cannulated and the liver was first perfused with heparined phosphate buffered saline “PBS” (+ penicillin, amphotericin B, streptomycin) to clear blood from the organ. The liver was then decellularized with 500 ml water containing 1% sodium dodecyl sulfate “SDS” and 1% Triton X for at least 6 hours each time using a pump with manual recirculation. In some livers we have tried to use some other specific enzymes. The organ was then washed with deionized water and then with PBS for about 24 hours. Successful decellularization was defined as the lack of nuclei or cytoplasmic staining using histological evaluation method with Hematoxilin Eosin. The decellularizated liver was then recellularized with regenerative cells. Results: This research work allowed us to obtain a bioartificial rat liver. In the process of decellularization and recellularization of the rat’s liver we have improved in some way the current technology and methods using a simple and effective apparatus for perfusion. An organ or tissue generated by this method could be transplanted to the rat model of chronic hepatitis. In certain instances, a decellularized organ may be recellularized with cells in vivo (e.g., after the organ could be transplanted into an individual). Engineering of a transplantable liver would be a permanent alternative to donor liver transplant. Conclusions: These results provide a proof of principle for the generation of a transplantable liver graft as a potential treatment for liver disease.