Introduction: Based on our previous findings, the treatment of stem cells alone or in combination with thyroid hormone (T 3 ) and mild exercise could effectively reduce the risk of stroke damage in young mice. However, it is unclear whether this treatment is effective in aged or middle-aged mice. Therefore, this study designed to assess whether combination of Bone Marrow Stromal Cells (BMSCs) with T 3 and mild treadmill exercise can decrease stroke complications in middle-aged mice. Methods: Under laser Doppler flowmetry monitoring, transient focal cerebral ischemia was produced by right Middle Cerebral Artery Occlusion (MCAO) for 45 min followed by 7 days of reperfusion in middle-aged mice. BMSCs (1×10 5 ) were injected into the right cerebral ventricle 24 h after MCAO, followed by daily injection of triiodothyronine (T 3 ) (20 μg/100 g/d SC) and 6 days of running on a treadmill. Infarct size, neurological function, apoptotic cells and expression levels of Glial Fibrillary Acidic Protein (GFAP) were evaluated 1 week after stroke. Results: Post-ischemic treatment with BMSCs or with T 3 and or mild treadmill exercise alone or in combination did not significantly change neurological function, infarct size, and apoptotic cells 7 days after ischemia in middle-aged mice (P>0.05). However, the expression of GFAP significantly reduced after treatment with BMSCs and or T 3 (P<0.01). Conclusion: Our findings indicate that post-stroke treatment BMSCs with exercise and thyroid hormone cannot reverse neuronal damage 7 days after ischemia in middle-aged mice. These findings further support that age is an important variable in stroke treatment

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This study was planned to continue our previous study to assess effect of combination therapy bone marrow stromal cells (BMSCs) with exercise (EX) and triiodothyronine (T 3 ) on stroke-induced astrogliosis in mice. Stroke subjected-mice were divided into five monotherapy groups including sham, control, BMSCs, EX and T 3 ; and three combination therapy groups including BMSCs + EX, BMSCs + T 3 and BMSCs + EX + T 3 . Astrogliosis was assessed in ipsilateral hemisphere at day 7 after MCAO.Combination therapy BMSCs with EX and T 3 could significantly decrease stroke-induced astrogliosis.However, monotherapy with BMSCs or EX also improved changes of glial fibrillary acidic protein (GFAP)positive cells following stroke. Combination therapy BMSCs with EX and T 3 didn't have any added effect on astrogliosis compared to monotherapy with BMSCs or EX. With comparing the present findings with the results of neurobehavioral functioning in our earlier study, it seems that decrease of astrogliosis could be helpful for stroke recovery.

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