Co-culturing of the Hepatocytes with the Endothelial and Stellate Cells within Decellularized Scaffolds by Sodium Lauryl Ester Sulfate- Approach to Tissue Constructs for Future Transplantation

Abstract

Background: Liver transplantation is the gold standard treatment for end-stage liver failure, but the scarcity of organ donors is the main limiting factor for performing liver transplant surgery.

Objective: The objective was to evaluate hepatocytes’ phenotype and functionalities after co-culturing with endothelial (HUVEC) and stellate cells (LX2) in the decellularized liver.

Methods: The livers were decellularized with 1% sodium lauryl ester sulfate (SLES). Cell removal and preservation of extracellular matrix (ECM) ultrastructure were studied by staining, scanning electron, and Raman confocal microscopy. The cell viability was evaluated by MTT, and the functions of cells were assessed on a decellularized scaffold with/without co-culturing with HUVEC and LX2 cell lines. The results were then compared to cells with the same condition on collagen scaffolds.

Results: The data confirmed that SLES prevented the destruction of the liver ECM ultrastructure along with nuclear material removal. Raman spectra confirmed DNA and cell debris removal. The decellularized liver was suitable for cell survival, but the proliferation rate was lower than those cultured in collagen. The tests showed that the function of individual cells on the decellularized scaffold was better than that in collagen scaffolds. Co-culturing with HUVEC and LX2 cell lines did not improve hepatocyte functions.

Conclusion: As a biocompatible scaffold, co-culturing hepatocytes with endothelial and stellate cells within the decellularized liver improved liver-specific functions.