The ex vivo productionofmature neutrophils depends on the sourceofCD34+ human hematopoietic progenitors
we propose a two-stage procedure for pre-expanding hematopoietic progenitor cells and subsequent ex vivo differentiation for potential granulocyte transfusion. We compared three distinct donor sources, UCB, BM live donor, and BM deceased donor, and tested two expansion different expansion protocols. Donor sources were evaluated based on proliferation during expansion and differentiation phases, neutrophil cell yield, and function. We show that cell for cell, UCB is the most proliferative and give the greatest neutrophil yield. Further, our ex vivo-generated neutrophils of all three donor sources have an enhanced propensity for opsonin-recognizing functions relative to primary neutrophils, which may influence future treatment. Overall design: Single-cell RNA-seq library preparation and sequencing: Single live cells were counted and processed using Honeycomb Single Cell RNA Sequencing platform HIVE. The samples were prepared for library preparation according to the manufacturer's protocol, and were then sequenced on an Illumina NextSeq500 sequencer.