Bone marrow-derived mononuclear cells (Postgrad Inst of Medical Education & Research)

Stem cells were harvested from autologous bone marrow obtained using a posterior superior iliac spine approach, under local anesthesia. Approximately 150 mL of bone marrow was harvested. Mononuclear cells (MNCs) were separated by ultracentrifugation after layering on density-gradient medium (Ficoll-Hypaque), washed with phosphate-buffered saline, and resuspended in heparinized normal saline (final volume: 5 mL). Aliquots were then taken for total nucleated cell count, MNC count, viability testing, using trypan blue, CD 34+ cell count, determined by flow cytometry, and sterility testing.