Isolation of porcine monocyte population: a simple and efficient method

Area of Science:

• Immunology
• Veterinary Science
• Cell Biology

Background:

• Monocytes are key immune cells involved in inflammatory responses.
• Established methods for human monocyte isolation contrast with limited protocols for porcine monocytes.
• Porcine monocytes are valuable for immunological research, but their isolation is challenging.

Purpose of the Study:

• To establish an efficient and cost-effective method for preparing and culturing high-purity porcine monocytes.
• To adapt and combine existing human monocyte isolation protocols for porcine applications.
• To achieve a reasonable yield of viable porcine monocytes suitable for experimental use.

Main Methods:

• Harvesting peripheral blood from pigs and treating with EDTA.
• Isolating peripheral blood mononuclear cells via density gradient centrifugation (Bicoll gradient).
• Culturing monocytes on serum-treated flasks, followed by rinsing and tapping; constant cooling was employed to enhance yield.
• Evaluating monocyte purity using CD14 expression via flow cytometry.

Main Results:

• The developed method achieved a high purity of porcine monocytes, averaging 92.6% (± 3.06%).
• The procedure is cost-effective and does not require specialized equipment.
• A reliable and reproducible method for preparing and culturing porcine monocytes was established.

Conclusions:

• A novel, cost-effective protocol for isolating and culturing porcine monocytes with high purity has been successfully established.
• This method offers a valuable tool for immunological research involving pigs.
• The technique is accessible and practical for laboratories without specialized equipment.