The production of large 'signalling competent' myeloid cells from circulating CD34+ cells in neonatal blood.
A method is described for the production of large myeloid cells, expressing functional formylated peptide receptors. CD34+ cells were isolated from neonatal cord blood by a two stage cell sorting method. Inclusion of SCF, together with IL-3, GM-CSF or both cytokines stimulated growth of these cells over 14 day period. The resultant cells, which ranged from 30 microns to 100 microns in size, were maintained in culture for up to 5 weeks, during which time the cell population increasingly displayed myeloid characteristics, including expression of formylated peptide receptors, phagocytosis and oxidase activation. These large cells had a functioning Ca2+ signalling system in response to the chemotactic peptide, f-Met-Leu-Phe. The large size of the cells enabled the rise in cytosolic free Ca2+ which resulted from either transmembrane influx of extracellular Ca2+ and release of Ca2+ from intracellular stores to be visualised. These large myeloid cells thus provide a model system for investigating the spatial characteristics of Ca2+ signalling by formylated peptide receptors on human myeloid cells.
Roberts
G M
GM
Department of Surgery, University of Wales College of Medicine, Health Park, Cardiff, UK.
Hoy
T
T
Hallett
M B
MB
D015703:Antigens, CD / Q000032:analysis*; D018952:Antigens, CD34; D002118:Calcium / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D015398:Signal Transduction
D015703:Antigens, CD; D018952:Antigens, CD34; D002118:Calcium