New method for constructing extracellular matrix bioscaffold derived from human cirrhotic liver tissues

Abstract

Objective To develop a novel decellularized extracellular matrix (ECM) bioscaffold derived from human cirrhotic liver tissues and construct a 3-dimensional cell culture system using this bioscaffold material. Methods Clinical samples of cirrhotic liver tissues were collected from patients undergoing liver transplantation for hepatic cirrhosis. The cirrhotic liver tissues were cleansed, cut into small tissue blocks (5 mm×5 mm×5 mm), and agitated in deionized water and detergents to obtain decellularized cirrhotic liver scaffold. Human primary human hepatocytes (PHHs) were seeded into the prepared ECM scaffold and cultured continuously for 9 d. Histological studies of the seeded cells were performed to assess the cell engraftment, and qPCR was used to detect the expression of the liver-specific genes in the cultured cells. Results The cirrhotic liver tissue became transparent after decellularization, and HE staining and scanning electron microscopy showed no residual nuclei in the bioscaffold. Residual DNA identification showed that the scaffold material had a residue DNA level less than 10 ng/mg of the tissue. After 9 d of continuous culture in the ECM scaffold, the PHHs showed good engraftment as shown by HE staining; the results of qPCR showed that the cells cultured in this 3D culture system expressed higher levels of liver-specific genes as compared with the cells in conventional 2D culture. Conclusion We successfully construct decellularized ECM bioscaffold derived from human cirrhotic liver tissues and confirmed the feasibility of 3D cell culture using this new bioscaffold material.

Keywords

• cirrhosis
• biological scaffold
• extracellular matrix