Peripheral blood-derived CD34+ cells (University Medical Centre Ljubljana)

CD34+ cells from peripheral blood were mobilized by granulocyte colony-stimulating factor and collected via apheresis. After 5 days, a full blood count and peripheral blood CD34+ cell count were performed. Peripheral blood stem cells were then collected with the Amicus cell separator. A magnetic cell separator was used for the immunomagnetic-positive selection of CD34+ cells. The collected cells were washed, to remove platelets, and then sensitized with mouse monoclonal anti-CD34 antibodies, and incubated with immunomagnetic beads coated with polyclonal sheep anti-mouse antibodies. The bead/CD34+ cell rosettes were separated from other cells and CD34+ cells were released from the Dynabeads using an octapeptide with an affinity for anti-CD34 antibodies. After immunomagnetic selection, cells were assessed for viability using methylene blue and reassessed for viability 2 hours thereafter, before intracoronary injection.