Human hepatocytes: isolation, culture, and quality procedures

Area of Science:

• Hepatology
• Cell Biology
• Biotechnology

Background:

• Isolated human liver cells are increasingly vital for research in liver diseases, drug development, and therapeutic strategies.
• High-quality cell isolation necessitates a robust logistical network involving surgeons, biologists, and technicians.

Purpose of the Study:

• To present a standardized protocol for isolating human liver cells from normal resected liver tissue.
• To discuss essential clinical networks, quality control measures, and the impact of 3D extracellular matrix on cultured cells.

Main Methods:

• Isolation of human liver cells from normal resected liver tissue.
• Establishment of a collaborative network for cell acquisition and processing.
• Culturing cells in 3D extracellular matrix to assess preservation of hepatocyte polarity.

Main Results:

• A reliable protocol for human liver cell isolation from resected tissue is described.
• The study emphasizes the role of a clinical network and quality controls for maintaining cell viability.
• 3D extracellular matrix culture preserves hepatocyte epithelial polarity and bile canaliculi formation, inhibiting epithelial-to-mesenchymal transitions seen in 2D cultures.

Conclusions:

• The presented protocol enables the isolation of high-quality human liver cells for R&D applications.
• Effective collaboration and stringent quality control are crucial for successful human liver cell culture.
• Utilizing 3D extracellular matrix is a promising strategy to maintain the in vivo-like phenotype of cultured hepatocytes.