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Fractionation of mammalian bone marrow by counterflow centrifugation‐elutriation using a continuous albumin gradient: analysis of granulocyte‐macrophage colony‐forming units
Author(s) -
Jemionek John F.,
MacVittie Thomas J.,
Byrne Patrick J.,
Schein Philip S.,
Walden Dane A.
Publication year - 1982
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.1365-2141.1982.tb01916.x
Subject(s) - elutriation , fractionation , bone marrow , centrifugation , granulocyte , differential centrifugation , albumin , biology , progenitor cell , cell , population , microbiology and biotechnology , immunology , haematopoiesis , andrology , chromatography , chemistry , biochemistry , stem cell , medicine , organic chemistry , environmental health
Bone marrow cells from murine, canine, primate and human donors were fractionated by counterflow centrifugation‐elutriation (CCE) using a continuous albumin gradient. Fractionation of 9 × 10 8 nucleated bone marrow cells (without prior removal of the erythrocytes) can be done in less than 1.5 h with 92% nucleated cell recovery, depending on donor species. Murine bone marrow cell recovery after fractionation averaged 52% whereas cell recoveries for canine, primate and human donors averaged 85%, 92% and 89%, respectively. The fractions were evaluated for total nucleated cell counts and granulocyte‐macrophage colony‐forming units (GM‐CFC). Each species studied presented a unique profile for nucleated cell recovery and associated GM‐CFC activity. These profile variations may suggest significant differences in the modes of GM‐CFC expression in the individual species that may depend on cell‐to‐cell interactions. Although CCE is unable to isolate a single population responsible for GM‐CFC activity, CCE does permit a rapid, reproducible fractionation of large numbers of cells with minimal manipulation of sample. This makes the isolated sample ideal for further purification of progenitor cell populations.