Expression Of Four Kinds Of MiRNA In Neural Stem Cells And Their Differentiation As Well As Motor Neurons

Background:BMSCs are one of the most researched non-neural tissue-derived adult neural stem cell. At present, the molecular mechanism of BMSCs differentiating into neurons was still unclear. Both miR-124 and miR-128 are abundant in the nervous system and they have better specificity. In this study, we detect the changes and effects of miR-124 and miR-128 in the process of bone marrow stromal cells (BMSCs) differentiating into neural cells.Objective:To detect the changes and effects of miR-124 and miR-128 in the process of bone marrow stromal cells (BMSCs) differentiating into neural cells.Methods:BMSCs were isolated and cultured with complete bone marrow stromal method. The cells were purified according to the ability of adherence. Then BMSCs of passage 3(abbreviated to P3) were induced into neural cells with induced medium. The morphologic features and growth status of BMSCs and differentiated BMSCs were observed by invert microscope. Real-time PCR was used to measure expression of miR-124 and miR-128 in the BMSCs and BMSCs after induction.Results:The level of miR-124 in differentiated BMSCs was 0.051-fold than that in BMSCs (P＜0.05). And the level of miR-128 in differentiated BMSCs was 0.070-fold than that in BMSCs (P＜0.05).Conclusions:miR-124 and miR-128 play an important role in the process of bone marrow stromal cells differentiating into neurons. Background:Neural stem cells are a group of self-renewing, self-proliferation and pluripotent cells. NSCs can be induced into cholinergic neurons; Hox is a key factor of the development of motor neurons in vivo, while the miR-126 can regulate and control Hoxa9, in this study we investigate that whether miR-126 also play a similar role in the process of SCSCs'development. miR-31 is an embryonic stem cell-specific miRNA. In our experiment, the SCSCs and spinal cord motor neurons were collected, and then the expression of miR-126 and miR-31 in the motor neurons and neural stem cell differentiation were detected to investigate their effects in neural stem cells during the neuron-oriented differentiation process.Objective:To detect the changes and effects of miR-126 and miR-31 in the motor neurons and in neural stem cells during the neuron-oriented differentiation process.Methods:Neural stem cells were collected from gestational age 17d of embryonic spinal cord tissue in rats, and then cultured and passaged. Nestin,the neuroepithelial stem proteins of the 3rd generation of Neural Stem Cells was detected by Immunofluorescence test.Then, the inducing factor RA, Shh, NT-3 and BDNF was used to induce the neural stem cells to differentiate. Two weeks after differentiation, the expression of choline acetyltransferase, ChAT was detected with immuno fluorescence。In another experiment, we isolated and purified embryonic spinal cord motor neurons by immunomagnetic beads method. Finally the ABI's TaqMan MicroRNA Assays real-time PCR technology was used to observe the changes and effects of miR-126 and miR-31 in the motor neurons and in neural stem cells during the neuron-oriented differentiation process.Results:The level of miR-126 in SCSCs was 0.142-fold than that in cholinergic neurons derived from SCSCs.The level of miR-31 in SCSCs was 0.118-fold than that in cholinergic neurons derived from SCSCs.The level of miR-126 in cholinergic neurons was 0.002-fold than that in motor neurons.The level of miR-31 in cholinergic neurons was 56.444-fold than that in motor neurons.The level of Hoxa9 in cholinergic neurons was 0.169-fold than that in motor neurons. Shh was expressed in the MNs, but can not be detected in cholinergic neurons by real-time PCR.Conclusions:The expression of miR-31 shows the difference between cells derived from SCSCs (cholinergic neurons) and desired cells (MNs); miR-126 and miR-31 play an important role in the differentiation process of NSCs.