Study Design Experimental animal study. Results Administration of CM from BMSC reduced apoptosis and inflammation at the site of injury in a rat model of SCI (p<0.05). Motor, sensory, locomotor, and sensorimotor performances were significantly improved in rats that received BMSC-CM after SCI. Conclusions Intrathecal administration of BMSC-CM improved recovery in a rat model of SCI. Keywords: Conditioned media, Spinal cord injuries, Breast milk, Stem cells Introduction Spinal cord injury (SCI) is a serious neurological condition that impacts a patients ability to function . SCI is a result of the primary injury, which occurs as a direct consequence of trauma and the secondary injury, which occurs following the onset of reactive processes such as inflammation, ischemia, free radical production, apoptosis, and necrosis in the spinal cord . SCI may lead to unpleasant complications, including various neurological problems such as the loss of sensory and motor function, autonomic nervous system dysfunction, pain, and ulcerous lesions . These issues underscore the need to develop better strategies for the treatment of SCI . SNS-032 (BMS-387032) With regard to these pathophysiological features, diverse treatment approaches have been suggested for SNS-032 (BMS-387032) SCI . Previous studies have mainly focused on controlling and limiting the mechanisms involved with protecting the nerves against secondary damage in the affected area . Replacing lost neurons and promoting neuronal regeneration using stem cells and their derivatives, such as stem cell-conditioned medium (CM), has been extensively studied [7,8]. Breast milk is usually a valuable and noninvasive source of heterogeneous stem cells. As a result, it has received considerable attention in the field of regenerative TLR9 medicine . A portion of the breast milk expresses markers for mesenchymal stem cells (MSCs) . Interestingly, a SNS-032 (BMS-387032) subpopulation of nestin-positive cells has also been reported in breast milk cells . Nestin is a marker for neural stem cells. Differentiation of this easily-available source of stem cells into a neural cell lineage has been previously reported . Although positive outcomes have been reported using stem cell therapy, there are still some disadvantages associated with these cells. For instance, the administration of stem cells may provoke an immune response or the formation of tumors . Using cell-derived CM is an effective approach that may overcome these problems associated with stem cell administration . Recent studies have revealed the neuroprotective effects of MSC-derived CM . It has also been reported that CM from MSCs may improve neuronal survival and promote neurite extension . However, there have been no studies conducted around the administration of breast milk stem cell (BMSC)-CM for SCI treatment. For this study, we examined the protective effects of BMSC-CM in a rat model of SCI. Materials and Methods 1. Experimental design Adult male Sprague-Dawley rats (250C280 g) were divided randomly into the following groups (n=6/group). (1) Control group: laminectomy was performed; (2) SCI/Sham group: Dulbeccos Modified Eagle Medium (DMEM)/F12 culture medium was injected intrathecally into the injured site after the induction of SCI; and (3) BMSC-CM group: BMSC-CM was injected intrathecally into the site of injury after the induction of SCI induction. This studys protocol was reviewed and approved by the university (REC.1393.998). 2. Isolation of cells from breast milk sample and characterization Breast milk was collected from healthy, lactating women who had given their consent prior to collecting samples. The samples were immediately transferred to the laboratory, diluted with an equal level of sterile phosphate-buffered saline (PBS), and centrifuged at.