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Inverse relationship between megakaryocyte buoyant density and maturity
Author(s) -
Jackson Carl W.,
Steward Shirley A.,
Brown L. Keith,
Look A. Thomas
Publication year - 1986
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1986.tb07571.x
Subject(s) - megakaryocyte , percoll , platelet , trypan blue , biology , microbiology and biotechnology , cell , centrifugation , immunology , haematopoiesis , biochemistry , stem cell
S ummary . We examined the relationship between rat megakaryocyte buoyant density and maturation stage in continuous Percoll density gradients. An average of 88% of megakaryocytes had buoyant densities <1–054 g/ml. There was an inverse relationship between megakaryocyte buoyant density and maturation. Morphologically mature forms comprised 90% of the megakaryocytes with buoyant densities of 1.030–1.033 g/ml. In contrast, immature morphology was present in three‐quarters of megakaryocytes with buoyant densities of 1.042–1.046 g/ml. These morphological findings were confirmed by [ 3 H]thymidine labelling studies. Cell viability assessed by trypan blue exclusion was highest among more dense megakaryocytes of which the majority were immature. The lowest trypan blue exclusion was found in the less dense, predominantly mature megakaryocytes indicating that these cells are more susceptible to membrane damage during marrow suspension. Megakaryocyte DNA content distributions and platelet antigen levels, determined by two‐colour flow cytometry, were also related to megakaryocyte density; the more dense megakaryocytes showed an approximately two‐fold higher proportion of 8N cells and less platelet antibody binding than did less dense megakaryocytes. These studies suggest that megakaryocytes can be fractionated according to their buoyant densities into immature and mature populations suitable for molecular studies of differentiation.