Long-term maintenance of functional primary human hepatocytes using small molecules

Area of Science:

• Hepatology
• Drug Metabolism
• Cell Culture

Background:

• Primary human hepatocytes (PHHs) rapidly deteriorate in culture, limiting their use in drug discovery.
• Maintaining hepatic functions like albumin and urea production, glycogen storage, and cytochrome P450 (CYP) expression is crucial for PHH utility.

Purpose of the Study:

• To develop a method to preserve the function of PHHs during extended culture.
• To evaluate the efficacy of a novel small molecule inhibitor cocktail (YPAC) in maintaining hepatic functions and CYP enzyme activity.

Main Methods:

• Treatment of PHHs with a cocktail of four small molecule signaling inhibitors (YPAC).
• Assessment of hepatic functions including albumin and urea productivity, glycogen storage, and CYP expression over 40 days.
• Measurement of CYP1A2, CYP2B6, and CYP3A4 enzymatic activities and CYP3A4 inducibility.

Main Results:

• YPAC effectively maintained various hepatic functions of PHHs during culture.
• PHHs treated with YPAC retained enzymatic activities of CYP1A2, CYP2B6, and CYP3A4 for up to 40 days.
• CYP3A4 activity inducibility by rifampicin and phenobarbital was preserved in YPAC-treated PHHs.

Conclusions:

• The YPAC cocktail is a valuable tool for maintaining PHH viability and function in vitro.
• This approach significantly enhances the utility of PHHs for drug discovery and toxicological studies.
• Extended culture of functional PHHs using YPAC can improve the accuracy of preclinical drug evaluation.