Megakaryocytes separation in homogeneous classes by unit gravity sedimentation: physico-chemical, ultrastructural and cytophotometric characterizations

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Abstract
Separation by velocity sedimentation at unit gravity according to the STAPUT system was applied to a population of rabbit megakaryocytes previously enriched by density gradient centrifugation. By this means, 80,000-100,000 megakaryocytes with 100% purity were collected in 8 fractions according to size for a sedimentation velocity of 52-30 mm/h. DNA-Feulgen cytophotometric measurements show significant correlation between megakaryocyte size and ploidy. The study of the 8 purified fractions is of particular interest because it reflects megakaryopoiesis evolution. The different stages of megakaryocyte maturation of each fraction were analyzed by transmission and scanning electron microscopy and were correlated to ploidy level. Thrombopoietic megakaryocytes with grape-like appearance were found in ploidy fractions 8n to 128n. Cytophotometric determinations of nucleohistones revealed several populations.