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Experimental Study of Mild Hypothermia and Neural Stem Cell Transplantation for Ischemic Nerve Dysfunction

Author: ChiFeng
Tutor: XuYingHui
School: Dalian Medical University
Course: Neurosurgery
Keywords: Neural stem cells MCAO Mild hypothermia Transplant
CLC: R651
Type: Master's thesis
Year: 2008
Downloads: 62
Quote: 0
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Abstract


Neural stem cells in terms of the research in recent years has become the hot neural science research field of stem cells separation, cultivation, purification and identification technology has been relatively mature, neural stem cell replacement treatment of nervous system diseases also gradually toward clinical application stage, but specific transplantation ways transplant conditions and transplantation time window has not yet formed a unified norms and standards of the experimental model of middle cerebral artery ischemia (MCAO) intracerebral transplantation of neural stem cells under conditions of mild hypothermia, observed under conditions of mild hypothermia transplant neural stem cells for the treatment of ischemic stroke effect, explore cell transplantation in patients with stroke and reliable alternative to treatment in a way to transplant, transplant conditions and transplantation time window, and confirmed the feasibility of the neural stem cell replacement therapy sex. Objective: To investigate the rat embryonic neural stem cell transplantation in the treatment of focal cerebral ischemia reperfusion injury in optimal conditions and efficiency. : From 13.5 days of gestation in Sprague-Dawley (SD) rats fetal rat cerebral cortex isolated neural stem cells, containing of bFGF, EGF, N2 and B27 factor DMEM/F12 joint purified neural stem cells cultured and Immunofluorescence was used to identify; selection the 270 ± 20gSD adult rats, rat MCAO model to take improved Koizumi suture method; induced Asia under low temperature conditions, select the third generation, the vigorous growth of neural stem cell spheres made of single cell suspension of rat model of intracerebral transplantation, three days prior to transplantation of neural stem cells to do the BrdU labeling and detection behavior, neurological function and brain were analyzed by immunohistochemistry 35 days after transplantation. Results: from the progestin 13.5 days in the fetal rat cerebral cortex successfully isolated neural stem cells, added bFGF, EGF, N2 and B27 factor DMEM/F12 (1:1) in vitro culture medium capable of stable growth, Amplification of neural stem cells, Nestin staining was positive 14 days after differentiation NeuN, GFAP and GalC staining positive results prove that the cultured cells into neural stem cells; 7 days after rat MCAO model was successfully the brain specimens visible obvious ischemic malacia forming and location constant, indicating the success of the model-making; 35 days after cell transplantation, compared with the negative control group, positive control group and the experimental group of mild hypothermia neurological score in statistically significant differences in transplant sites were detected BrdU-positive neural stem cells, the immune group results found that transplanted stem cells differentiation and the formation of NeuN positive neurons. Conclusion: The application contains of bFGF, EGF, N2 and B27 combined with the medium, from 13.5 days of gestation fetal rat cerebral cortex neural stem cells isolated and cultured and used in further experimental studies; produce a higher rate of success, good repeatability and large rat MCAO experimental animal models; mild hypothermia combined transplantation of neural stem cell survival and differentiation in the rat cerebral ischemic lesions localized effect of transplantation in the behavioral and histological better than a simple neural stem cell transplantation.

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