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The Effects of Muscle-derived Stem Cells on Neurons Damage Induced by Oxidative Stress in a Co-culture System

Author: LiuShiQiong
Tutor: MeiXiFan
School: Liaoning Medical
Course: Surgery
Keywords: Muscle-derived stem cells Cortical neurons The Millicell plug cell culture dishes ( the Transwell small room ) Brain derived neurotrophic factor Nerve - like cells Bcl-2 Bax Neuron-specific enolase Poly-L-lysine T- butyl hydrogen peroxide
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 59
Quote: 0
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Abstract


Purpose, looking for a new primary cell culture method can effectively improve the neurons and muscle-derived stem cells in quantity and quality to provide a sound basis for later experiments. 2, muscle-derived stem cells and neurons co-cultured, the observation of muscle-derived stem cells differentiate into neuron-like cells. 3, the muscle-derived stem cells and oxidative stress in neurons co-cultured to observe the conditions of muscle-derived stem cells co-cultured neurons have the protection of oxidative stress injury. Method 1, using the hybrid enzyme digestion were isolated and cultured neurons and muscle-derived stem cells, the cytarabine and differential adhesion purified, inverted microscope cell morphology was observed, cell count, growth curve, analysis of muscle-derived stem cell proliferation. Desmin (desmin) and neural neuron specific enolase (NSE) immunohistochemical methods were used to identify the muscle-derived stem cells and neurons. 2, muscle-derived stem cells with oxidative stress (injury group) or without oxidative stress-treated neurons (not injury group) co-culture, the establishment of a control group (stem cell group), muscle-derived stem cells cultured alone during collection of three groups of cell culture medium. Cultured for 6 days after its the NSE identification, muscle-derived stem cell differentiation differences observed between the different groups, and three sets of culture medium ELISA assay to explore the potential causes. 3, oxidative stress neurons and muscle-derived stem cells co-cultured, the control group for the oxidative damage to neurons cultured alone. Survival of neurons in each group at different time points difference detected through Hoster33258 and flow cytometry, and related genes by RT-PCR analysis to explore this difference underlying reasons. Results after mixed enzyme digestion, the successful high-purity, high activity of rat cortical neurons (NSE-gt; 95%) and muscle-derived stem cells Desmin (positive rate of Gt; 92%). 2, cultured for 6 days after the detected damage group and non-injury group were part of the myogenic differentiation of stem cells for the expression of NSE neuron-like cells, the former high differentiation of the latter, the stem cell group there was no cell differentiation into neuron-like cells. The first two days of injury group BDNF secretion is higher than the non-injured group and blank control group (P lt; 0.05), while the first two days of the injury group BDNF secretion is also higher than the secretion of BDNF 4 and 6 days (P lt; 0.01), also higher than the other groups at all time points secretion (P lt; 0.01). Injury group and non-injury group than the control group in the fourth day of the sixth day secretion in. BDNF secretion in the culture medium of the three time points injury group and non-injury group and blank control group were higher than the MDSCs group (P lt; 0.001). In the case of oxidative stress injury, injury (muscle-derived stem cells co-cultured) neuron survival is much higher than the control group (not myogenic, found through Hoster33258 and flow cytometry stem cells co-cultured), RT-PCR found damaged neurons of BCL-2 expression levels related with the former. Conclusion 1, NSE, and Desmin cells immunochemical confirmed high purity cortical neurons and muscle-derived stem cells, mixed enzyme digestion is a simple and effective in vitro cultured neurons and muscle-derived stem cells were cultured in vitro. In co-culture conditions, rat cortical neurons can induce muscle-derived stem cells into neuron-like cells, and neurons secrete BDNF is a vital link. Muscle-derived stem cells can also secrete BDNF. 3, the next day, the fourth day, the sixth day of three time points, neuron survival of the injury group were higher than the control group, muscle-derived stem cells can produce a protective effect on damaged neurons, Bcl-2 gene upregulation of Bax down is one of the reasons.

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