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The Use of Mannitol for Bone Marrow Mesenchymal Stem Cells Transplantation

Author: LiuJinPing
Tutor: MoXueAn
School: Guangxi Medical University
Course: Neurology
Keywords: Rats Bone marrow mesenchymal stem cells Cell culture Vascular Dementia Animal models Morris water maze Cognitive function Mannitol Transplant Brain - derived neurotrophic factor
CLC: R749.13
Type: Master's thesis
Year: 2011
Downloads: 54
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Abstract

Charge-quality stem cells of the first part of rat bone marrow separation, culture and identification 【purpose】 explore rat bone marrow mesenchymal stem cells (Bone marrow mesenchymal stem cells, BMSCs) of in vitro separation, purification, passages, and identification, for transplantation in the treatment provided function stable BMSCs. To take childhood SD rat femur under sterile conditions 【Methods】 whole bone marrow culture adherent method combined with cell separation and purification of BMSCs. After the cell growth of about 80% -90% of the bottom of the bottle is covered with the plastic culture control with 0.25% trypsin digestion and passage in the 30-60s. Primary and passaged cell morphology was observed under inverted phase contrast microscope. Identification of the BMSCs cultured and expanded by osteogenic differentiation method and flow cytometry. [Results] rat BMSCs in vitro the separable, culture and amplification. Isolated and cultured BMSCs morphology fusiform adherent growth of fibroblast-like cells were passaged cell morphology tends to be uniform. Induce BMSCs can differentiate into osteoblasts, Alizarin Red S visible calcium nodules positive alkaline phosphatase, visible in pink positive granules in the cytoplasm. The BMSCs of surface markers by flow cytometry, almost uniform expression of CD44 and CD90. CD44 and CD90-positive rate of 89.4% and 99.2%, respectively, while the CD34-positive rate was 1.82%. [Conclusion] whole bone marrow binding to cells adherent method can be successful to establish the rat BMSCs vitro isolation and culture system, and the cultured BMSCs high purity, vitality stable traits. BMSCs induced to osteoblast differentiation, with multiple differentiation potential. The second part of vascular dementia rat model preparation [Objective] To investigate the preparation the ideal vascular dementia (Vascular dementia, VD) animal model to provide reliable VD animal models for the next experiment. 【Method】 interval of three days has bilateral common carotid artery ligation prepared the VD rats model using the Morris water maze cognitive function in rats. [Results] The postoperative 4w, the survival rate of rats up to 52.17%. Cognitive function test results show that a significant cognitive dysfunction, compared with the sham group, the VD model rats. [Conclusion] VD model of the improved preparation of 2-VO, a high success rate, reproducible, stable symptoms of dementia, can be prepared by the ideal of VD model to be used for infrastructure experimental study. The third part mannitol pretreatment underwent bone marrow mesenchymal stem cell transplantation efficacy stayed rat model of vascular disease observed 【Objective】 To observe the of intravenous mannitol pretreatment bone marrow stromal stem cells (Bone marrow mesenchymal stem cells BMSCs) Vascular dementia vascular dementia (VD) rat cognitive function test brain organization of brain-derived neurotrophic factor (Brain-derived neurotrophic factor, BDNF) expression levels. 【Methods】 The intravenous injection of mannitol (1.5g.kg-1 body weight) pretreatment 10-30mins, 1 × 106/mL BMSCs1ml implanted through the tail vein of VD rats, 4w, using water maze test, enzyme-linked immunosorbent assay (Enzyme-linked immunosorbent assay, ELISA) method detects the VD rats with cognitive function as well as brain tissue BDNF content change. 【Results】 compared with medium control group of BMSCs transplantation in the treatment group and mannitol pretreatment BMSCs transplantation in the treatment group were improved cognitive function in rats model rats escape latency (P lt; 0.05), the platform quadrant District residence time (P lt; 0.05); with BMSCs transplantation in the treatment group compared to, to improve the the mannitol pretreatment BMSCs transplantation treatment group cognitive function in rats, the escape latency shortening (P lt; 0.01), the the platform quadrant District residence time (P lt; 0.05). Hippocampus, frontal cortex BDNF content: compared with medium control group of BMSCs transplantation in the treatment group and mannitol pretreatment the BMSCs transplantation treatment group in the rat brain BDNF expression levels increased (P lt; 0.01); with the BMSCs transplant treatment comparison group, the mannitol pretreatment BMSCs transplantation in the treatment of group rat brain BDNF expression increased (P lt; 0.01). [Conclusion] mannitol therapy after intravenous pretreatment BMSCs VD rat models, improved cognitive function, the cortex and hippocampus of BDNF expression levels than separate intravenous BMSCs treatment mouse model better than a single intravenous injection of BMSCs treatment the increased rat model.

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