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Ping G.,Hebei Provincial Laboratory of Brain Aging and Cognitive Neuroscience | Ping G.,Hebei Medical University | Zhang Z.-X.,Hebei Provincial Laboratory of Brain Aging and Cognitive Neuroscience | Wang Y.-Y.,Hebei Provincial Laboratory of Brain Aging and Cognitive Neuroscience | And 5 more authors.
Journal of Clinical Rehabilitative Tissue Engineering Research | Year: 2010

BACKGROUND: Good results have been achieved by the transplantation of bone marrow stromal cells (BMSCs) and neural stem cells (NSCs) separately into the brain of Parkinson's disease (PD) model rat, while the transplantation of conditioned medium of BMSCs associated with NSCs has not been reported. OBJECTIVE: To observe the effects of intracephalic transplantation of NSCs combined with BMSC conditioned medium on the behavior and cognition of PD model rats, and compare with NSC transplantation alone. METHODS: Following 24 hours of culture using Neurobasal+B27, BMSCs at 3-6 passages were harvested and the supernatant was centrifuged, i.e., BMSC conditioned medium. Following in vitro culture, rat NSCs were labeled by BrdU. Of 55 rats, 8 rats were randomly selected as a normal group. The remaining rats were used to establish PD models by injection of 6-hydroxydopamine (6-OHDA) in the lateral substantia nigra and ventral tegmental area using a stereotactic apparatus. Thirty-two models were successfully established and randomly assigned to three groups. Two coordinate points at the right corpus striatum were selected as a transplantation point. Simple NSC group was injected with NSC suspension (5 μL) at each point. Combination group was infused with the mixture of NSC suspension + BMSC conditioned medium (5 μL). Model group did not receive any liquid. Behavior changes were observed in PD rats. Cognition function was assessed in rats using a Morris water maze test. Tyrosine hydroxylase protein expression in the substantia nigra and BrdU expression in the transplanted region were detected utilizing immunohistochemistry. RESULTS AND CONCLUSION: Compared with model group, the rotation times of rats in simple NSC group, combination group decreased significantly from 1st to 8th weeks after transplantation (P < 0.01). The escape latency of rats in simple NSC group and combination group was significantly decreased (P < 0.05). The crossing times, percentage of swimming distance and swimming time in the platform quadrant in the Morris water maze test were increased after transplantation (P < 0.05). No significant difference was found in each index between the later two groups (P > 0.05). Expression of tyrosine hydroxylase-immunoreactive cells and nerve fiber expression were deleted in the rat substantia nigra undergoing 6-OHDA in each group at 8 weeks following transplantation. BruU-positive cells were determined in the transplanted regions in the simple NSC and combination groups; most located near to needle channel, and some migrated along the corpus callosum. Results indicated that transplantation of NSCs combined with BMSC conditioned medium can improve rotational behavior and cognitive ability of PD rats, which is similar to corpus striatum transplantation of simple NSCs.

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