Studies of bone marrow cells have shown that they can restore neurologic function when transplanted into animals with ischemic brain injury. Recently, our laboratories have isolated a specific multi-potent stem cell population from human bone marrow that is capable of differentiating into neurons, astrocytes, and oligodendroglia in vitro under appropriate culture conditions. We postulate that these bone marrow-derived stem cells can provide an autologous source of donor cells for transplantation and repair in conditions of ischemic brain injury and stroke.
In Specific Aim 1 we will determine the neural phenotypes generated from bone marrow-derived human stem cells when transplanted into the brain of spontaneously hypertensive rats. Human stem cells will be transduced to express eGFP to label the donor cells. Bone marrow-derived stem cells will then be transplanted into the striatum, hippocampal formation, or sensorimotor cortex. At various time periods after transplantation, brain tissue from host rats will be studied to identify presence of eGFP labeled cells. eGFP labeled cells will also be examined to determine whether they develop into neurons, glial cells, and/or oligodendroglia. Site-specific differentiation will also be evaluated to determine whether grafted neurons also co-express neurotransmitter markers that are characteristic of neurons that found in the striatum, hippocampus, and cortex.
In Specific Aim 2 we will determine the efficacy of transplanted bone-marrow stem cells in ameliorating neurologic deficits associated with ischemic brain injury. The middle cerebral artery will be occluded unilaterally by ligation distal to the branching striatal vessels. This occlusion produces a discrete ischemic lesion of the cortex and results in permanent sensory and motor deficits in the forelimb contralateral to the side of injury. One week after the ischemic event animals will receive transplants of bone marrow-derived stem cells into regions of the neocortex surrounding the area of injury. In preliminary studies we have found that these transplants are capable of ameliorating sensorimotor deficits. The extent of the functional recovery will be assessed over time using a battery of behavioral tests. Also as part of Specific Aim 2 we will determine the time window after ischemic brain injury for stem cell transplantation that will result in an amelioration of neurologic deficits. In these studies ischemic rats will receive transplants at various time periods after ischemic injury and assessed functionally over time using a defined set of sensory and motor tests.
In Specific Aim 3 we will investigate possible mechanisms underlying bone marrow-graft induced recovery of neurologic function. Possible mechanisms include trophic effects on neural stem cells of the host brain, the induction of growth factor release by host brain cells, and the reorganization of host fiber connections. The results of these studies will provide information regarding the efficacy of using specific bone marrow-derived stem cells as an autologous source of cells for transplantation in ischemic brain injury, and information regarding possible mechanisms of graft-induced neural plasticity.
