Cellular expansion of hepatocellular carcinoma organoids using decellularized liver scaffolds.

BACKGROUND

A decellularized liver scaffold (DLS) is a three-dimensional acellular extracellular matrix created by removing cellular components from liver tissue. Hepatocellular carcinoma (HCC) organoids represent a useful experimental model.

METHODS

HCC organoids from patient-derived xenografts (PDX), liver organoids, and HepG2 cells were expanded by cultivation within a murine DLS.

RESULTS

HCC and liver organoids were generated from HCC PDX and human liver tissues, respectively. Expression levels of hepatocyte paraffin 1 (HepPar1), epithelial cell adhesion molecule (EpCAM), alpha-fetoprotein, keratin-7, and keratin-19 were detected in normal liver tissue, HCC tissue, HCC PDX, and HCC organoids. Fifteen murine DLSs were created, and the complete absence of liver cells was confirmed histologically by Masson trichrome and periodic acid-Schiff staining. Culture of HCC organoids in the DLS resulted in the expansion of numerous HCC cells scattered throughout the DLS framework. Keratin-7 expression was abundant, indicating widely dispersed progenitor cells. In contrast, cultivation of HepG2 cells within the DLS resulted in sparse cell distribution throughout the scaffold. Human hepatocyte organoids could not be cultivated successfully within the murine DLS framework.

CONCLUSIONS

The DLS collagen framework facilitates the proliferation of HCC organoids. Thus, cultivating HCC organoids within a DLS appears to represent an effective method for rapid cell expansion, although it was ineffective for HepG2 cells and liver organoids. Further studies are required to validate the biological and oncological equivalence between seeded HCC organoids and those rapidly expanded within a DLS.