The aim of this study was to evaluate the effect of 5-fluorouracil mobilized bone marrow regenerative cells (BMRCs) transplantation on brain injury following focal cerebral ischemia and to explore the mechanisms.Male Sprague-Dawley rats were subjected to middle cerebral artery occlusion (MACO) for 120 minutes, followed by intravenous administration of DAPI-labeled 1×10(7) 5-fluorouracil mobilized BMRCs at 24 h post MACO. Infarct volumes, neurological deficit score, angiogenesis and cytokine expression were evaluated at specific time point after cell transplantation.Comparable number of BMRCs and bone marrow cells (BMCs) were found in the infarcted area at Day 3 and Day 14 post MACO. Significant decreased infarcted size and neurological deficit score were found in the animals receiving BMRCs. The microvessel density was significantly increased after BMRCs transplantation. Moreover, the expression of vessel endothelial growth factor (VEGF) was increased significantly after BMRCs transplantation.Intravenous 5-fluorouracil mobilized BMRCs were neuroprotective following MACO and might be considered as therapeutic choice in the treatment of transient focal cerebral ischemia.