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Isolation of normal human megakaryocytes
Author(s) -
Sitar Giammaria
Publication year - 1984
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.1365-2141.1984.tb03993.x
Subject(s) - centrifugation , megakaryocyte , suspension (topology) , bone marrow , population , chromatography , sedimentation , suspension culture , chemistry , cell culture , biology , immunology , materials science , pathology , progenitor cell , microbiology and biotechnology , medicine , stem cell , mathematics , genetics , paleontology , environmental health , homotopy , sediment , pure mathematics
S ummary . This paper reports a simple procedure for obtaining human megakaryocytes with a high purification and high recovery yield. Bone marrow cells, obtained from surgically removed ribs, were separated by a two‐step procedure. Initially, a single cell suspension was enriched in megakaryocytes by equilibrium density centrifugation, the low density cell fraction was subsequently layered over a shallow continuous albumin gradient in a glass sedimentation chamber. Megakaryocytes averaged 0.03 ± 0.02% of all nucleated cells in the starting marrow cell suspension, after this procedure an average 80 ± 15% of the initial megakaryocyte population was recovered with a purity of 94 ± 4%. Previous methods, based upon the use of a two‐step procedure, are reviewed. The theory of velocity sedimentation is discussed with regard to the differences in the methodology used, which account for the different results I obtained.