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Identification of mRNA for PDGF B‐chain in human megakaryocytes isolated using a novel immunomagnetic separation method
Author(s) -
Gladwin A. M.,
Carrier M. J.,
Beesley J. E.,
Lelchuk R.,
Hancock V.,
Martin J. F.
Publication year - 1990
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.1365-2141.1990.tb06364.x
Subject(s) - megakaryocyte , riboprobe , microbiology and biotechnology , platelet derived growth factor receptor , immunomagnetic separation , biology , messenger rna , platelet , platelet derived growth factor , megakaryocytopoiesis , growth factor , haematopoiesis , chemistry , in situ hybridization , gene , immunology , biochemistry , stem cell , receptor
Summary A rapid and simple method has been developed for the separation of pure populations of intact human megakaryocytes from whole bone marrow. Megakaryocytes were specifically recognized using monoclonal antibodies coupled to magnetizable articles. Cells labelled with the magnetizable probe were separated from unlabelled cells by introduction of a magnetic field. The technique yields megakaryocyte suspensions with a purity of > 98%. Electron microscope examination showed that the ultrastructure of the isolated megakaryocytes was well preserved. Using this method of cell purification, we have investigated expression of the gene for platelet‐derived growth factor (PDGF). RNA was isolated from cells harvested from either ribs or posterior iliac crest. The RNA was spotted onto nitrocellulose, and then hybridized using a c‐sis riboprobe specific for PDGF B chain mRNA. We demonstrate that mRNA for PDGF B‐chain is identifiable in samples of 50000 cells. We conclude that PDGF is synthesized by the megakaryocyte.