Differential Analysis of Rat Bone Marrow by Flow Cytometry

Abstract:

A procedure for rat bone marrow differential analysis using flow cytometry and commercially available monoclonal antibodies is described. The method uses a combination of the differential expression of leucocyte common antigen (CD45) on different cell lineages and the expression of transferrin receptor (CD71). This is coupled with the side scatter measurement (SSC) for morphological complexity and nucleic acid staining with LDS-751 for the separation of all nucleated cells from mature red blood cells. The relative number of erythroid, myeloid and lymphoid cells as well as the M:E ratio was generated by the flow cytometric method with acceptable reproducibility and a good agreement with microscopic analysis. There were no significant differences in mean values of the three cell populations between males and females or between bone marrow samples collected from left and right femurs. The flow cytometric analysis of bone marrow provides a semi-automated and rapid method that is suitable for the evaluation of rat bone marrow in the preclinical toxicity investigations.