Isolation of human megakaryocytes by density centrifugation and counterflow centrigual elutriation

Abstract

Density centrifugation and counterflow centrifugal elutriation were utilized to prepare enriched fractions of megakaryocytes from human bone marrow aspirates. This separation method enriched megakaryocytes in initial marrow aspirates by 116- to 463-fold. Approximately 63% of megakaryocytes were recovered from the initial samples, composing 18.7% of the nucleated cells in the final preparations. Mean megakaryocyte diameters of 51.6 micron and 33.8 micron were obtained from fixed and unfixed cellular specimens, respectively. Smaller platelet glycoprotein-positive mononuclear cells with a mean diameter of 20.5 micron were found in the highest concentrations in this final fraction. These cells presumably represent immature megakaryocytic forms. Counterflow centrifugal elutriation provides a means of isolating enriched populations of marrow megakaryocytes. This accessibility to viable populations of human megakaryocytes will allow additional investigation of the terminal events of megakaryocyte development.