Flow cytometric evaluation of bone marrow differentials in rats with pharmacologically induced hematologic abnormalities

• Published in: Cytometry (New York, N.Y.) Vol. 32; no. 1; pp. 18 - 27
• Main Authors: Criswell, K. A., Bleavins, M. R., Zielinski, D., Zandee, J. C., Walsh, K. M.
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.05.1998
• Subjects: 2′7′ dichlorofluorescin; Animals; Bone Marrow Cells; Bone Marrow Examination - methods; Cell Count; cyclophosphamide; Cyclophosphamide - pharmacology; erythropoietin; Erythropoietin - pharmacology; Female; Flow Cytometry - methods; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; granulocyte–macrophage stimulating factor; Hematologic Diseases - chemically induced; Hematologic Diseases - pathology; hematology; Hyperplasia; Male; peroxidase; Rats; Rats, Wistar; recombinant murine stem cell factor; Stem Cell Factor - pharmacology
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/(SICI)1097-0320(19980501)32:1<18::AID-CYTO3>3.0.CO;2-B

Previously, flow cytometric determination of peroxidase activity, cell size, and reactivity to lymphocyte antibodies were used to produce bone marrow differentials in untreated rats. In the present study, abnormal hematologic profiles were induced with erythropoietin (EPO), recombinant murine stem cell factor (rm‐SCF), granulocyte–macrophage stimulating factor (GM‐CSF), and cyclophosphamide (CP). Manual and flow cytometric data showed comparable levels of erythroid and myeloid hyperplasia in EPO‐ and rm‐SCF/GM‐CSF‐treated animals, respectively. In CP‐treated animals, flow cytometric data revealed significant decreases in cellularity at concentrations of CP ≥ 5 mg/kg. In contrast, 20 mg/kg CP were necessary to induce microscopically apparent hypoplasia in histologic bone sections, showing that the automated methodology was a more sensitive indicator of bone marrow hypocellularity than was the more conventional manual method. Megakaryocyte counts were consistently higher by flow cytometer than by manual counts performed on cytocentrifuge preparations made from the same cell suspensions but were similar to megakaryocyte counts performed on histologic sections of femur, indicating that the automated methodology produced a more accurate reflection of true megakaryocyte numbers. Induction of hematologic abnormalities in the present study showed that manual bone marrow differentials can be replaced with the more efficient and reliable flow cytometric method in most preclinical toxicology studies. Cytometry 32:18–27, 1998. © 1998 Wiley‐Liss, Inc.