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Megakaryocytes separation in homogeneous classes by unit gravity sedimentation: physico‐chemical, ultrastructural and cytophotometric characterizations
Author(s) -
Dupont H.,
Dupont M. A.,
Bricaud H.,
Boisseau M. R.
Publication year - 1984
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1111/j.1768-322x.1984.tb00231.x
Subject(s) - biology , megakaryocyte , ultrastructure , sedimentation , sedimentation coefficient , population , centrifugation , ploidy , differential centrifugation , specific gravity , homogeneous , microbiology and biotechnology , anatomy , progenitor cell , mineralogy , genetics , biochemistry , chemistry , stem cell , sediment , sociology , gene , enzyme , physics , thermodynamics , paleontology , demography
Separation by velocity sedimentation at unit gravity according to the STAPUT system of Miller and Phillips was applied to a population of rabbit megakaryocytes previously enriched by density gradient centrifugation. By this means, 80,000 to 100,000 megakaryocytes with 100% purity were collected in eight fractions according to size for a sedimentation velocity of 52 to 30 mm/hr. DNA‐Feulgen cytophotometric measurements show significant correlation between megakaryocyte size and ploidy. The study of the eight purified fractions is of particular interest because it reflects megakaryopoiesis evolution. The different stages of megakaryocyte maturation of each fraction were analysed 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.