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

W. L. Smith, Lisa A Tell, M. B. Kabbur, L. Gage, James S Cullor

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)37-42
Number of pages6
JournalComparative Clinical Pathology
Volume8
Issue number1
StatePublished - 1998

Keywords

  • Asian elephant
  • Flow cytometry
  • Heterophil isolation
  • Oxidative product formation
  • Phagocytosis

ASJC Scopus subject areas

  • Anatomy
  • Pathology and Forensic Medicine

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