Polymorphonuclear leukocytes isolated from umbilical cord blood as a useful research tool to study adherence to cell monolayers

Janhavi Sharma; Prerna Rastogi; Michael H Creer; Jane McHowat

doi:10.1016/j.jim.2009.09.008

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Polymorphonuclear leukocytes isolated from umbilical cord blood as a useful research tool to study adherence to cell monolayers

Journal article

Peer reviewed

Polymorphonuclear leukocytes isolated from umbilical cord blood as a useful research tool to study adherence to cell monolayers

Janhavi Sharma, Prerna Rastogi, Michael H Creer and Jane McHowat

Journal of immunological methods, Vol.351(1-2), pp.30-35

12/31/2009

DOI: 10.1016/j.jim.2009.09.008

PMCID: PMC2783264

PMID: 19800347

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Abstract

One of the initial steps in the inflammatory process involves the adherence and transmigration of circulating polymorphonuclear leukocytes (PMN) across the endothelial cell monolayer. One of the main constituents of the neutrophil phagosome that contributes to bacterial killing is myeloperoxidase (MPO) which can be measured spectrophotometrically, using hydrogen peroxide as a substrate, and hence can be used as an index to quantify neutrophil adherence. To evaluate whether PMN isolated from umbilical cord blood could be used for in vitro experiments to monitor neutrophil adherence, we compared the adherence to confluent endothelial and epithelial cell monolayers using PMN isolated from umbilical cord and adult peripheral blood. The extent of PMN adherence was assessed by measuring MPO activity. In initial experiments, we isolated PMN from umbilical cord and adult peripheral blood and measured MPO activity with respect to cell number and assay incubation times. Our data demonstrate that PMN obtained from either source had similar MPO activity and similar adherence to endothelial or epithelial cells. In conclusion, our data suggest that umbilical cord blood is a suitable source of leukocytes to examine PMN adherence in the setting of inflammation in a variety of disease processes.

Neutrophils - cytology

Peroxidase - immunology

Respiratory Mucosa - cytology

Endothelial Cells - metabolism

Humans

Cells, Cultured

Neutrophils - immunology

Cell Separation - methods

Fetal Blood - cytology

Cell Adhesion - immunology

Endothelial Cells - immunology

Pregnancy

Apoptosis - immunology

Endothelial Cells - cytology

Respiratory Mucosa - immunology

Annexin A5

Adult

Female

Respiratory Mucosa - metabolism

Neutrophils - metabolism

Peroxidase - metabolism

Details

Title: Subtitle: Polymorphonuclear leukocytes isolated from umbilical cord blood as a useful research tool to study adherence to cell monolayers
Creators: Janhavi Sharma - Department of Pathology, Saint Louis University School of Medicine, 1402 S. grand Blvd., St Louis, Missouri 63104, USA
Prerna Rastogi
Michael H Creer
Jane McHowat
Resource Type: Journal article
Publication Details: Journal of immunological methods, Vol.351(1-2), pp.30-35
DOI: 10.1016/j.jim.2009.09.008
PMID: 19800347
PMCID: PMC2783264
NLM abbreviation: J Immunol Methods
ISSN: 0022-1759
eISSN: 1872-7905
Publisher: Netherlands
Grant note: DK66119 / NIDDK NIH HHS R01 DK066119 / NIDDK NIH HHS R01 DK066119-05 / NIDDK NIH HHS
Language: English
Date published: 12/31/2009
Academic Unit: Pathology
Record Identifier: 9984047715202771

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