FLOW-CYTOMETRY OF DNA CONTENT IN HUMAN-BONE MARROW - A CRITICAL REAPPRAISAL

Abstract

Because cytokinetic studies of the human bone marrow aspirate as a prognostic factor and as a monitor of drug perturbation [produced by antineoplastic agents on the cell cycle distribution of normal and leukemic bone marrow] are frequently inconsistent, reproducibility of DNA distribution was measured by flow cytometry of DNA content in patients with morphologically normal bone marrow. In 15 patients correlation was noted between DNA distributions simultaneously obtained on right and left iliac crest bone marrow aspirates (r [correlation coefficient] = 0.588), although considerable variation in individuals was encountered. Much better reproducibility (r = 0.879) was achieved using bilateral core biopsy of bone marrow in these same patients. In 60 samples, comparison of DNA distribution between bone marrow aspirate and simultaneously obtained biopsy revealed higher relative proportions of S and G2 + M phase cells in biopsies (P < 0.001), suggesting peripheral blood contamination of aspirate material. Brisk shaking of biopsy specimens in saline expelled a representative sample in the supernatant that could be subjected to simultaneous cytomorphological and cytokinetic analysis. To improve reproducibility of DNA content determinations in normal human bone marrow, bone marrow biopsy should be utilized.