Ted Robert
Due to their short life span native Poly-Morphonuclear Neutrophils (PMNs) are difficult to genetically manipulate. We have developed an optimal
culture protocol for the ex vivo production of neutrophils from CD34+ stem cells isolated from peripheral blood. This protocol produces a 70%
population of mature neutrophils with over 500-fold cell expansion. The resulting neutrophils display similar nuclear morphology and cell-surface
marker expression profiles as native PMNs, while mass spectrometry analysis indicates that cultured neutrophils possess a highly similar proteome
to PMNs. Additionally, stem cell- derived neutrophils also display an activatable respiratory burst and can produce Neutrophil Extracellular
Traps (NETs). We are currently further characterizing stem cell-derived neutrophils, assessing the efficiency at which they phagocytose pathogens,
degranulate and kill bacteria. Although numerous studies have produced neutrophils from CD34+ stem cells, as far as we are aware, this will
be the first-time stem cell-derived neutrophils have been fully characterised. We plan to utilise this culturing system, along with the expertise the
Toye lab has in genetic manipulation of early haematopoietic progenitors using lentivirus, to examine numerous aspects of neutrophil cell biology.
In close collaboration with Dr Borko Amulic, we plan to focus on aspects that are difficult to study using neutrophil like cell lines, including NET
formation and degranulation, as well as neutrophil disorders and differentiation defects.
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