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Am J Physiol Cell Physiol (April 2, 2003). doi:10.1152/ajpcell.00035.2003
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Submitted on January 23, 2003
Accepted on March 25, 2003

A novel method to quantify the turnover and release of monocytes from the bone marrow using the thymidine analog, 5'-bromo-2'-deoxyuridine

Yukinobu Goto1, James C Hogg1, Tatsushi Suwa1, Kevin B Quinlan1, and Stephan F van Eeden1*

1 Department of Pathology, McDonald Research Laboratories and iCAPTURE Centre, University of British Columbia, Vancouver, British Columbia, Canada

* To whom correspondence should be addressed. E-mail: svaneeden{at}mrl.ubc.ca.

The present study was designed to develop methods to study the production and release of monocytes from the bone marrow (BM) using the thymidine analog, 5'-bromo-2'-deoxyuridine (BrdU). Dividing monocytes in BM were labeled with BrdU (MOBrdU) and their release into the blood and disappearance from the circulation were monitored using a double immunostaining method. The first MOBrdU appeared in the circulation 4 h after labeling with BrdU, peaked at 18 h when 34.3 ± 5.8% of monocytes were labeled. The calculated transit time of monocytes through BM was 38.1 ± 3.1 h in control rabbits with a half-life (T1/2) of 12.7 h. Instillation of Streptococcus pneumoniae into the lung accelerated the release of monocytes from BM (peak at 10 h) and shortened their BM transit time (27.1 ± 1.8 vs. 22.6 ± 0.6, vehicle vs. pneumonia, p<0.05). We conclude that this non-radioisotope method provides a novel way to monitor monocyte kinetics and confirmed previous reports that a focal pneumonia shortens monocyte marrow transit and increases their release into the circulation.




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