Comparison of methods for isolating primary hepatocytes from mini pigs

Area of Science:

• Hepatology
• Biomaterials Science
• Cell Biology

Background:

• Porcine hepatocyte isolation is vital for bioartificial liver devices and transplantation.
• Formulated collagenases (Serva) are used for tissue isolation.
• N-acetylcysteine (NAC) enhances human hepatocyte viability.

Purpose of the Study:

• To compare the efficacy of two Serva collagenase NB grades (NB 4 and NB 8) for porcine hepatocyte isolation.
• To evaluate the impact of N-acetylcysteine (NAC) on hepatocyte isolation when combined with Serva NB 8 collagenase.
• To assess hepatocyte viability, yield, and metabolic function post-isolation.

Main Methods:

• Porcine hepatocytes were isolated using the perfusion method from Bama mini pigs.
• Three groups were studied: Serva NB 4, Serva NB 8, and NB 8 + NAC.
• Hepatocyte viability, spheroid formation, cell yield, and metabolic function (albumin and urea synthesis) were assessed after 24-hour culture.

Main Results:

• The NB 8 + NAC group demonstrated superior hepatocyte viability (97.8±1.9%) compared to NB 8 (94.4±2.4%) and NB 4 (94.5±3.2%).
• The NB 8 + NAC group achieved a higher cell yield ((11.8±1.0)×10^9 cells) than NB 8 ((9.5±2.1)×10^9) and NB 4 ((9.1±1.1)×10^9).
• Superior albumin secretion ((425.8±35.3) ng/mL) and reduced hepatocyte injury were observed in the NB 8 + NAC group.

Conclusions:

• Formulated collagenase Serva NB 8 in combination with NAC significantly enhances porcine hepatocyte isolation.
• This optimized method results in higher yields of viable porcine hepatocytes with improved metabolic function.
• The findings support the use of Serva NB 8 collagenase and NAC for improved hepatocyte isolation in liver research and therapeutic applications.