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Transplantation of Umbilical Cord Mesenchymal Stem Cells into Cisterna Magna for Cerebral Hypoxic-ischemic Rat Model

Author: ZhongYanYan
Tutor: HuangYaLing
School: Huazhong University of Science and Technology
Course: Diseases of the nervous system in children
Keywords: Newborn rats Hypoxic-ischemic encephalopathy Animal models Neuroethology Umbilical cord mesenchymal stem cells Cisternals Behavior
CLC: R722.1
Type: Master's thesis
Year: 2011
Downloads: 5
Quote: 0
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Abstract


Objective: To explore to intrauterine hypoxic ischemic encephalopathy model making and evaluation methods. Methods: 20 days pregnant pregnant SD rats were randomly divided into experimental group (n = 12) and control group (n = 3). Experimental group ligation pregnant rat uterine artery for 15 minutes, natural childbirth, born to the control group of mice intrauterine global cerebral hypoxic-ischemic injury model mice; mice born to normal pregnant rats. Recording the number of each set of experimental mice, the mortality was calculated. D1 ~ D16 weight monitoring, observation eyes open, expand time pinna, with a nervous reflex test suspension experimental mine experiments, reject prisoners reaction test, the turn stick cytometry experiments Morris water maze exercise of the experimental group and the control group evaluation and cognitive functions. Randomly taken from newborn pups do pathological observation. Results: 163 of the total number of fetal rat model group, 47.2% of the stillbirth rate, 1 week mortality rate of 6.9%. Weight gain of the experimental group than the control group slowly, eyes open and ear to start time delay compared with the control group. Part of the neural reflex delays compared with the control group; suspension experimental model group suspended time is significantly shorter than the control group (P lt; 0.05); the Scotch Miriam experimental model group residence time is significantly shorter than the control group (P lt; 0.05); Kuang wear grid frequency and stool grains the field experimental model mice was significantly less than the control group (P lt; 0.05), and while standing times compared to the control group, no significant difference; to reject prisoners reaction experimental model mice compared to the control group is difficult crawl; Morris water maze experiments mice get hidden platform position significantly longer time than the control group, the short residence time in the target quadrant than the control group. HE staining observed by light microscopy, and inflammatory cells in the cerebral cortex and hippocampus of model group gathered neurons was significantly reduced degeneration and necrosis of neurons and glial cells reactive hyperplasia. Conclusion: ligation of the uterine artery of pregnant rats preparation intrauterine hypoxia ischemia animal model can successfully mimic human HIE model. Objective: To explore the umbilical cord-derived mesenchymal stem cells (UCMSCs) transplantation in the treatment of developing rats with hypoxic-ischemic encephalopathy (HIE) the effectiveness of sequelae. Methods: Human umbilical cord tissue after collagenase and trypsin digestion, separation, cultured UCMSCs, the DII marked, prepared and to mark UCMSCs. The uterine artery ligation prepared model of cerebral ischemia and hypoxia was born 20 days (D20) mice as the experimental group, were randomly divided into stem cell group (n = 39) and PBS group (n = 39); stem cell group the cisterna magna puncture the injection the DII marked the UCMSCs, PBS group cisterna magna puncture PBS. 1,6 weeks after transplantation, immunohistochemistry was used to detect DII, neuron-specific enolase (NSE), Nestin (Nestin), glial fibrillary acidic protein (GFAP). Experiment by hanging, Scotch cytometry experiments and Morris water maze behavioral assessment, record the score. Natural childbirth, normal pregnant rats born to mice as a control group, and the same period in the control and experimental groups. And for statistical analysis. Results: 1 week, 6 weeks after transplantation of the stem cells found DII, NSE, Nestin and GFAP-positive cells in the hippocampus and cortical motor area. Suspension experiments, two weeks after the transplant, the stem cell group compared to the PBS group, the control group suspension lasted for a long time, the three difference was statistically significant (p lt; 0.05). Turn stick cytometry experiments, three weeks after the transplant, the stem cell group compared with the PBS group, the control group in turn rod instrument on the long residence time, the three difference was statistically significant (p lt; 0.05). Morris water maze test, the average incubation period of the stem cell group, PBS group, the control group difference was statistically significant (p lt; 0.05), at different times average incubation period the difference was statistically significant (p lt; 0.05) between the groups and the time No significant interaction (p = 0.0939). Space exploration, experimental single-factor analysis of variance showed 60s stem cell group, PBS group, the control group swimming time in the target quadrant (quadrant 3), three no statistical difference (P gt; 0.05). The stem cell group, PBS group, the control group, the first time through the platform location, three no statistical difference (P gt; 0.05). Conclusion: Human umbilical cord mesenchymal stem cells transplanted the cisterna magna to hypoxic-ischemic brain injury in rats model can migrate to the hippocampus and cortical motor area, and differentiate into neural stem cells, neurons and astrocytes; exogenous stem cell transplantation to promote the recovery of brain function to some extent.

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CLC: > Medicine, health > Pediatrics > Newborns, premature children disease > Neonatal disease
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