Long-term survival and functional maintenance of hepatocytes by using a microfabricated cell array

Area of Science:

• Hepatology and Regenerative Medicine
• Biotechnology and Bioengineering

Background:

• Primary hepatocytes lose function rapidly in standard culture, limiting their use in research and drug testing.
• Developing stable, functional hepatocyte models is crucial for understanding liver biology and disease.

Purpose of the Study:

• To develop a microfabricated cell array for long-term culture of hepatocyte spheroids.
• To assess the viability and functional maintenance of hepatocytes within these spheroids.
• To evaluate the utility of this model for liver disease research and preclinical drug development.

Main Methods:

• Harvesting fresh hepatocytes from Wistar rats.
• Coculturing hepatocytes with feeder cells in a microfabricated array to form spheroids.
• Monitoring spheroid formation and stability over 42 days.
• Assessing key hepatocyte functions: albumin secretion and cytochrome P450 (CYP) metabolic activity.

Main Results:

• Hepatocyte spheroids demonstrated long-term viability and maintained their structural integrity for 42 days.
• Cocultured spheroids retained essential cellular functions, including albumin secretion and CYP metabolic activity, throughout the 42-day period.
• The microfabricated cell array facilitated the stable formation and maintenance of functional hepatocyte spheroids.

Conclusions:

• The novel coculture method using microfabricated arrays enables the long-term maintenance of functional hepatocyte spheroids.
• This advanced model holds significant potential for advancing liver disease research and improving the early prediction of hepatocyte toxicity in drug development.
• The sustained viability and function of these spheroids offer a more reliable platform for in vitro studies compared to traditional hepatocyte cultures.