Mouse liver cell culture. I. Hepatocyte isolation

Area of Science:

• Hepatology
• Cell Biology
• Biochemistry

Background:

• Accurate isolation of viable mouse liver cells is crucial for various biological studies.
• Existing methods for liver cell isolation often require optimization for yield and cell integrity.

Purpose of the Study:

• To describe and optimize a two-step perfusion method for isolating mouse liver cells.
• To compare variations of existing rat liver cell isolation procedures for mouse models.
• To assess cell yield, viability, and morphological integrity throughout the isolation process.

Main Methods:

• A two-step perfusion technique using calcium and magnesium-free Hanks' salt solution followed by a collagenase-containing medium.
• Quantitative comparison of procedural variations for cell yield and viability.
• Light and electron microscopy to evaluate liver cell morphology at different isolation stages.

Main Results:

• The optimal method involves hepatic portal vein perfusion, yielding an average of 2.3 x 10^6 viable liver cells/g body weight.
• An optimal collagenase concentration of 100 U/mL was determined.
• Hanks' solution caused reversible cell injury, which was reversed by collagenase perfusion, preserving cell morphology.

Conclusions:

• The described two-step perfusion method provides an efficient and reliable technique for isolating high-yield, viable mouse hepatocytes.
• The optimized procedure minimizes cell damage and preserves the morphological integrity of isolated liver cells.
• This method is suitable for applications requiring pure, functional mouse liver cells.