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  • Analysis of rat bone marrow by flow cytometry following in vivo exposure to cyclohexanone oxime or daunomycin HCl.

Analysis of rat bone marrow by flow cytometry following in vivo exposure to cyclohexanone oxime or daunomycin HCl.

Toxicology and applied pharmacology (2002-12-04)
Shelli J Schomaker, Frances A S Clemo, David E Amacher
ABSTRACT

The purpose of these studies was to evaluate bone marrow from male CD rats following exposure to known hematotoxins using flow cytometry (FC) and a monoclonal antibody to the cell surface antigen CD71. Rats were treated with either CHO (300 mg/kg for 10 days) or DAUN (10 mg/kg for 1 day). Control groups received the appropriate vehicle. Half of the animals from each group were euthanized at the end of the dosing schedule and the remaining animals were euthanized after a recovery period. Hematology analyses were completed prior to the onset of each study and on the day of necropsy. Marrow was isolated from the tibia, stained with R-phycoerythrin-conjugated mouse anti-rat CD71 (transferrin receptor on proliferating cells) monoclonal antibody, and then analyzed by FC for myeloid:erythroid (M:E) ratios. FC determinations of myeloid and erythroid population percentages and M:E ratios from untreated rats were confirmed by microscopic examination of marrow cytospins and selected flow cell sorts. M:E ratios for control animals determined by FC were not significantly different between the two studies (1.83 vs 1.89). CHO treatment caused a significant (p < 0.01) decrease in M:E ratios (0.96 for CHO vs 1.48 for control) at day 11 due to increased erythroid cells. M:E ratios were significantly increased (p < 0.05) with DAUN treatment at day 3 (5.07 for DAUN vs 1.70 for control) and corresponded to generalized depletion of all marrow cell lines, especially erythroid cells. After recovery, M:E ratios of CHO and DAUN rats were similar to controls. Hematological values corroborated changes in marrow myeloid and erythroid populations evaluated by this FC technique. Using FC and a monoclonal antibody to the cell surface antigen CD71, this study confirmed the reversible selective toxicity on myeloid and erythroid marrow populations following in vivo exposure to CHO or DAUN. This FC procedure provides a rapid, sensitive method for bone marrow analysis compared to conventional cytological examination.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cyclohexanone oxime, 97%