Bone marrow-derived mononuclear cells (Medical University of Vienna)

Bone marrow was aseptically harvested from the posterior iliac crest of each patient (300 ml), under general anesthesia, the day before therapy. Bone-marrow cells were anti-coagulated with 10 IU/ml heparin. After collection, the bone marrow was passed through two filters with different pore sizes (500 µm and 200 µm). Volume depletion and enrichment of bone-marrow-derived mononuclear cells were performed with the COBE SPECTRA[R]. Bone marrow was diluted with acid citrate dextrose solution A at a ratio of 1:10 to avoid coagulation during the step before the in vitro processing of the cells. Plasma pump rates were adjusted during processing to maintain the red blood cell plasma interface within the white blood cell collection channel. The proportions of the CD34+ cells were determined by fluorescence-activated cell sorting analysis.