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A rapid isolation of Asian elephant (Elephas maximus) blood heterophils on Percoll density gradients

A rapid isolation of Asian elephant (Elephas maximus) blood heterophils on Percoll density gradients This is the first report of a rapid technique for the isolation of elephant heterophils form ethylene diamine tetra-acetic acid (EDTA) anticoagulated whole blood using hetastarch sedimentation and Percoll discontinuous gradient centrifugation modified from equine neutrophil isolation techniques (Sedgwick et al. 1986; Pycock 1987). Heterophil purity and viability was greater than 98% and 99%, respectively. The integrity of the isolated heterophils was evaluated by light microscopy and flow cytometric forward (size of cell) and orthogonal light-scattering (granularity) properties. In addition, functional properties including phagocytosis and oxidative product formation were evaluated and compared to heterophils within a buffy coat (centrifuged blood) population. Light microscopic morphological features including toxic or pyknotic changes, general size, cellular membrane and cytoplasmic granule integrity of heterophils from peripheral blood, buffy coats and isolated heterophils were similar. Flow cytometric forward and orthogonal light-scattering properties were unchanged. No significant differences between isolated cells and heterophils within buffy coat preparations were evident for phagocytosis or oxidative product formation. Variation in percentage phagocytosis (66% to 86%) and mean channel fluorescence intensity (2288 to 3625) in isolated heterophils was observed among individual elephants. This may be attributed to functional heterogeneity of heterophils among elephants. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Comparative Clinical Pathology Springer Journals

A rapid isolation of Asian elephant (Elephas maximus) blood heterophils on Percoll density gradients

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Publisher
Springer Journals
Copyright
Copyright © 1998 by Springer-Verlag London Limited
Subject
Medicine & Public Health; Hematology; Oncology; Pathology
eISSN
1433-2973
DOI
10.1007/BF02628103
Publisher site
See Article on Publisher Site

Abstract

This is the first report of a rapid technique for the isolation of elephant heterophils form ethylene diamine tetra-acetic acid (EDTA) anticoagulated whole blood using hetastarch sedimentation and Percoll discontinuous gradient centrifugation modified from equine neutrophil isolation techniques (Sedgwick et al. 1986; Pycock 1987). Heterophil purity and viability was greater than 98% and 99%, respectively. The integrity of the isolated heterophils was evaluated by light microscopy and flow cytometric forward (size of cell) and orthogonal light-scattering (granularity) properties. In addition, functional properties including phagocytosis and oxidative product formation were evaluated and compared to heterophils within a buffy coat (centrifuged blood) population. Light microscopic morphological features including toxic or pyknotic changes, general size, cellular membrane and cytoplasmic granule integrity of heterophils from peripheral blood, buffy coats and isolated heterophils were similar. Flow cytometric forward and orthogonal light-scattering properties were unchanged. No significant differences between isolated cells and heterophils within buffy coat preparations were evident for phagocytosis or oxidative product formation. Variation in percentage phagocytosis (66% to 86%) and mean channel fluorescence intensity (2288 to 3625) in isolated heterophils was observed among individual elephants. This may be attributed to functional heterogeneity of heterophils among elephants.

Journal

Comparative Clinical Pathology – Springer Journals

Published: May 3, 2007

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