Effects Of Intravenous Administration Of Marrow Stromal Cells On Expression Of Brain-derived Neurotrophic Factor In Vascular Dementia Rat

PARTⅠIsolation,Cultivation And Identification Of Rat Bone Marrow-derived Mesenchymal Stem Cells【Objective】To investigate rat bone marrow mesenchymal stem cells (Mesenchymal stem cells, MSCs) in vitro isolation, purification, mass generation, identification, offer cell transplantation seed cells with stable function.【Method】sterile conditions, take young SD rat femur, combined with whole bone marrow culture adherent cell separation and purification of MSCs. When cells covered with plastic bottom 80% -90%, digest them with 0.25% trypsin digestion 30-60S then passage the cells.Use inverted phase contrast microscope to observate primary cells and passaged cells'morphology . Osteogenic differentiation of MSCs to identify MSCs after cell amplification.Combination of whole bone marrow culture and cell attachment way can successfully construct a rat MSCs isolation and culture system in vitro, and cultured MSCs with high purity, energy intensity, stable characters. MSCs have multiple differentiation potential, can be induced into osteoblasts. 【Results】Rat MSCs were isolated , cultured and amplified in vitro. MSCs were growed with adherence and exhibited a spindle-shaped fibroblastiv morphology.spindle shape, After passaged into the third generation MSCs'morphology become similar. Alizarin red staining showed the calcium mineralization positive.【Conclusion】Combination of whole bone marrow culture and cell attachment way can successfully construct a rat MSCs isolation and culture system in vitro, and cultured MSCs with high purity, energy intensity, stable characters. MSCs have multiple differentiation potential, can be induced into osteoblasts.PARTⅡEstablishing Vascular Dementia Model In Rats【Objective】To establish an animal model of vascular dementia (VD) in rats.【Methods】Vascular dementia was induced by clipping the bilateral common carotid arteries in rats at an interal of 7 days for each artery. Abilities to learning and memory of rats were tested by using the localization sailing test with Morris water maze.【Results】Survival rate of VD rates reach up to 82% by 30 days postoperatively. Compared with sham group, the created rat VD model showed significant alteration in leaning and memory function.【Conclusions】This model of VD proved to be reliable,reproducible, and had on stable status. It has the practicability and reliability and can be used in basic experiment study. PARTⅢA Observation On The Therapeutic Effect Of Transplantation Of Bone Marrow-derived Mesenchymal Stem Cells On The Vascular Dementia Rats【Objective】To observe change of the expression of Brain-derived neurotrophic factor in the brain of Vascular dementia rat after injecting Mesenchymal stem cells into intravenous.【Methods】Mesenchymal stem cells marked by Brdu were injected into intravenous of Vascular dementia rat.After 4 weeks, the recognition of VD rat were observe and change of BDNF in brain were detected by water maze and enzyme-linked immunosorbent assay .【Results】Comparing to model group,There are statistics discrepancy in both localization sailing test and space pilot test (P < 0.01) of rat receiving transplantion of Mesenchymal stem cells.The content of BDNF were improved in hippocampus, Cortex of frontal lobe and cerebrospinal fluid(0.524±0.382,0.879±0.410;0.150±s0.131,0.333±0.192;0.160±0.141,0.268±0.142).【Conclusion】After transplantion of Mesenchymal stem cells, space learning and memory of VD rat were improved and this improvement may concerne with increase of BDNF.