Preparation of a decellularized scaffold derived from human liver tissue

METHODS: A surgical specimen of the left lateral lobe of the liver was obtained from a patients with hepatic hemangioma. The decellularization process was performed by repeated freezing-thawing, sequential perfusion with 0.01% SDS, 0.1% SDS and 1% Triton X-100 through the portal vein, and sterilization with peracetic acid. L-02 cells were then engrafted onto the decellularized liver scaffold.

RESULTS: HE staining, DAPI staining and scanning electron microscopy all verified the absence of residual cellular components in the decellularized scaffold. The residual DNA content in the decellularized scaffolds was 25.3∓14.6 ng/mg (dry weight), which was less than 1% of the total DNA content in a fresh human liver. Immunohistochemistry demonstrated that type I and IV collagens, fibronectin and elastin were all retained in the scaffold. The engrafted L-02 cells survived well on the scaffold with active proliferation and expressed albumin and G6pc.

CONCLUSION: It is feasible to prepare decellularized scaffolds using surgical specimens of human liver, which can be a new approach to constructing a tissue-engineered liver for clinical purposes.

OBJECTIVE: To develop a method for preparing a decellularized scaffold based on human liver tissue.