Isolation of equine peripheral blood mononuclear cells using Percoll

Area of Science:

• Veterinary Immunology
• Cell Biology
• Hematology

Background:

• Accurate isolation of equine peripheral blood mononuclear cells is essential for immunological research.
• Previous protocols using 60% Percoll have shown suboptimal cell yields.

Purpose of the Study:

• To determine the optimal Percoll concentration for isolating equine peripheral blood mononuclear cells.
• To improve cell yield and purity for subsequent functional assays.

Main Methods:

• Equine peripheral blood was processed using density gradient centrifugation with varying Percoll concentrations.
• Cellular yield and purity were assessed at different Percoll concentrations.

Main Results:

• A 60% Percoll concentration resulted in poor cell yield.
• A 65% Percoll concentration significantly increased the yield of mononuclear cells.
• A 70% Percoll concentration was effective in retaining neutrophils while isolating mononuclear cells.

Conclusions:

• The optimal Percoll concentration for isolating equine mononuclear cells for lymphocyte and monocyte function studies is 65%.
• A 70% Percoll concentration can be utilized to isolate pure equine neutrophils.