The morphology and function of rabbit hepatocytes isolated using ethylenediaminetetraacetate

Area of Science:

• Hepatology
• Cell Isolation Techniques
• Organ Preservation

Background:

• Collagenase is the standard enzyme for hepatocyte isolation but has limitations.
• Hepatocyte isolation is crucial for transplantation and research.
• Developing alternative, effective isolation methods is essential.

Purpose of the Study:

• To evaluate ethylenediaminetetraacetate (EDTA) as an alternative to collagenase for rabbit hepatocyte isolation.
• To assess the preservation of isolated hepatocytes using University of Wisconsin (UW) solution and Dulbecco's modified eagle's medium (DMEM).
• To compare the morphological and functional viability of preserved hepatocytes.

Main Methods:

• Hepatocyte isolation using concentrated EDTA solution followed by Percoll gradient purification.
• Preservation of isolated hepatocytes at 4°C for up to 72 hours in UW solution or DMEM.
• Morphological assessment using Papanicolaou and PAS stains.
• Functional assessment including LDH release, water content, and 99m-technetium mebrofenin uptake.

Main Results:

• EDTA perfusion provided reproducible hepatocyte isolation with yields comparable to collagenase.
• Morphological studies showed pure hepatocytes with preserved architecture in UW solution up to 72 hours.
• Functional studies indicated significantly lower LDH release and water content, and higher 99m-technetium mebrofenin uptake in UW solution-preserved hepatocytes.

Conclusions:

• Concentrated EDTA solutions enable reproducible isolation of pure, viable rabbit hepatocytes.
• UW solution effectively preserves hepatocyte morphology and function at 4°C for up to 72 hours.
• EDTA-based isolation and UW solution preservation offer a promising method for hepatocyte research and transplantation.