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

Journal of immunological methods (2009-10-06)
Janhavi Sharma, Prerna Rastogi, Michael H Creer, Jane McHowat
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with ribonucleosides, deoxyribonucleosides and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture