Low back pain and subsequent disabilities are common. A lumbar spinal fusion procedure is an effective treatment with autologous bone grafts, but harvesting the bone from the iliac crest is associated with risks of complications. New treatments using stem cells together with osteoconductive and otesoinductive materials have made the procedure safer, but the inconsistency of the amount of stem cells harvested from bone marrow aspirate still remains to be solved. This study reports that the bone dusts, usually discarded as surgical wastes during transforaminal lumbar interbody fusion procedure (TLIF procedure), yielded cells which had the characteristics of mesenchymal stem cells (MSCs) in vitro. The cells were positive for the MSC markers and were able to differentiate in osteogenic and adipogenic directions. The cells grew robustly in an osteoconductive material, Bolheal (serum glue), and also proliferated well in culture medium supplemented with autologous serum. Therefore, the bone dust is a good candidate for the alternative source of stem cells other than bone marrow aspirate to increase the safety of the TLIF procedure.