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Difference in the Expression of Notch-1 Gene during Muscle-derived Stem Cells Repairing Spinal Cord Injury in Rats

Author: ZhangHe
Tutor: MeiXiFan
School: Liaoning Medical
Course: Surgery
Keywords: MDSCs NGF Nerve -like cells BrdU Notch pathway Notch1
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 31
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Abstract

Objective To investigate the muscle-derived stem cells (Muscle derived stem cells, MDSCs) Notch1 gene expression differences in repair rat spinal cord injury. Method 1, the machinery - mixed enzyme digestion were isolated and cultured rat MDSCs in desmin (Desmin) Immunocytochemistry identified as muscle-derived stem cells. NFG induced MDSCs using immunohistochemistry assay after induction of cell-specific markers expressed NSE. 2, according to the experimental requirements, select 120 rat samples, animal random number table method is divided into: group A: of NGF induction and BrdU labeling the MDSCs group (n = 30); Group: B uninduced BrdU labeling MDSCs group (n = 30); C: control group (n = 30) and Group D: blank control group (n = 30). A, B, C group sampling time is divided into one week, four weeks, eight weeks. Surgical methods: Make a spinal cord hemisection model. MDSCs cultured fluid after spinal cord injury using micro-syringe injection of NGF-induced differentiation and BrdU labeling to Group A, the B group was injected with the same amount without induction of differentiation but BrdU labeling MDSCs culture medium, C group were injected with the same dose of PBS fluid. 4 rats in each group at different time points in survival during the determination of the rats hind limb motor function (the BBB motor function score and the inclined plane test). The immunohistochemistry staining observed of MDSCs In rats survival, migration and differentiation circumstances. 6, RT-PCR and Western detection of muscle-derived stem cells after implantation, the expression of key genes in the Notch pathway. 1, after the machinery - mixed enzyme digestion and differential adherence successful cultivation of high purity, high activity of MDSCs, Immunocytochemistry results for Desmin () (positive rate gt; 80%). 2, the induction process, the good state of the cell growth, morphology was a gradual change, no neurospheres formed; the induced group 7d after nerve-like cells in cell body and neurites NSE stained strongly positive (37.29 ± 1.59)% without induction group cells positive cell rate is extremely low (4.26 ± 0.34)%, obvious differences between the two groups (P lt; 0.01); the induction group MDSCs samples detected the NSE expression levels higher than the non-induced group (P lt; 0.01). 3, the behavior observed results: Group A (induction group) 1 week after injury rated slightly higher than that of group B (non-induced group) but were higher than group C (control group), with time, the group differences gradually The obvious group A score significantly higher than group B, significantly better than the non-intervention group C. The results show that the muscle-derived stem cell transplantation in the treatment of spinal cord injury induced by NGF were significantly better than the muscle-derived stem cell transplantation alone. Histological results: observed after BrdU labeling the transplanted MDSCs in rats survival, migration and differentiation. Group A damage repair particularly evident in the number of cells than B group. The results showed that, after NGF after induction of MDSCs to repair spinal cord injury in rats better than MDSCs in alone. RT-PCR and Western detection results show: the Notch signaling pathway in Notch1 node gene mRNA and Notch1 protein showed a low expression in the rat spinal cord injury, the induction group and the non-induced group had significant differences (P lt; 0.05) . Conclusion 1, the experimental machinery - mixed enzyme digestion and cultured in vitro MDSCs primary cultured to obtain a high-purity, high activity the MDSCs, Desmin Immunocytochemistry identified proved. 2, the present study, the NGF of MDSCs induced in vitro to differentiate into neuron-like cells implanted in rat spinal cord injury at the treatment and achieved certain results, MDSCs as seed cells to repair spinal cord injury. 3 in rat spinal cord injury Notch signaling pathway Notch1mRNA and the Notch1 protein showed low expression Notch signaling molecules with low expression can promote MDSCs to differentiate into neuron-like cells.

Difference in the Expression of Notch-1 Gene during Muscle-derived Stem Cells Repairing Spinal Cord Injury in Rats

Abstract

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