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 journalArticle

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

Fingerprint

Blood Buffy Coat
Phagocytosis
Light
Hydroxyethyl Starch Derivatives
Cytoplasmic Granules
Diamines
Poisons
Centrifugation
Cell Size
Acetic Acid
Horses
Microscopy
Neutrophils
Fluorescence
Membranes
Percoll
Population

Keywords

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

ASJC Scopus subject areas

  • Anatomy
  • Pathology and Forensic Medicine

Cite this

A rapid isolation of Asian elephant (Elephas maximus) blood heterophils on percoll density gradients. / Smith, W. L.; Tell, Lisa A; Kabbur, M. B.; Gage, L.; Cullor, James S.

In: Comparative Clinical Pathology, Vol. 8, No. 1, 1998, p. 37-42.

Research output: Contribution to journalArticle

@article{0ab73ff2350b45ddb088351620da87d4,
title = "A rapid isolation of Asian elephant (Elephas maximus) blood heterophils on percoll density gradients",
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.",
keywords = "Asian elephant, Flow cytometry, Heterophil isolation, Oxidative product formation, Phagocytosis",
author = "Smith, {W. L.} and Tell, {Lisa A} and Kabbur, {M. B.} and L. Gage and Cullor, {James S}",
year = "1998",
language = "English (US)",
volume = "8",
pages = "37--42",
journal = "Comparative Clinical Pathology",
issn = "1618-5641",
publisher = "Springer London",
number = "1",

}

TY - JOUR

T1 - A rapid isolation of Asian elephant (Elephas maximus) blood heterophils on percoll density gradients

AU - Smith, W. L.

AU - Tell, Lisa A

AU - Kabbur, M. B.

AU - Gage, L.

AU - Cullor, James S

PY - 1998

Y1 - 1998

N2 - 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.

AB - 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.

KW - Asian elephant

KW - Flow cytometry

KW - Heterophil isolation

KW - Oxidative product formation

KW - Phagocytosis

UR - http://www.scopus.com/inward/record.url?scp=54749139247&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=54749139247&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:54749139247

VL - 8

SP - 37

EP - 42

JO - Comparative Clinical Pathology

JF - Comparative Clinical Pathology

SN - 1618-5641

IS - 1

ER -