Isolation and Chemiluminescent Properties of Ferret (Mustela putorius furo) Polymorphonuclear Cells

Abstract

ABSTRACT. Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formylMet-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets. KEY WORDS: chemiluminescence assay, ferret (Mustela putorius furo), polymorphonuclear cell.

Chemotactic properties and absence of the formyl peptide receptor in ferret (Mustela putorius furo) neutrophils

“…Equine neutrophils also do not demonstrate any chemotactic response to fMLF, and there is no inflammatory reaction when fMLF is injected intradermally, despite the presence of FPR (Brazil et al, 1998). Moreover, the authors previously reported that ferret PMNs do not generate superoxide in response to fMLF (Nakata et al, 2007). Taken together, ferret neutrophils appear to lack FPR and consequently do not have chemotaxis and superoxide generation activity through the FPR.…”

“…This study was approved by the Ethical Committee for Animal Experimentation, NUVERC. Isolation of PMNs was performed as previously described (Nakata et al, 2007). In brief, the whole blood was mixed with 2% dextran solution (dextran T500: Pharmacia, in saline), and then left for 20 min at room temperature.…”