Short-term Withdrawal Of BFGF And EGF Effects On The Proliferation And Differentiation Of Neural Stem Cells

Epidermal growth factor (EGF) is a peptide of53amino acid residues, and its internal conservative of the three disulfide bonds. EGF in the body’s various tissues and body fluids, widespread, and a variety of tissue cells has a strong mitogenic effect. The in vitro experiments show that EGF in the development of late to promote neiral precursor cell proliferation and differentiation. And promote differentiation of of collagen progenitor cells play a role.Fibroblast growth factor (FGF) is a155-268amino acid residues connected by small peptides.Active substance is a way to promote fibroblast growth. Basic fibroblast growth factor (FGF2) as the fibroblast growth factor family member of the play to promote neural stem cells in mitosis in early embryos and to maintain its survival, and glial cells and neurons, progenitor cells havepromote differentiation. In addition, neural stem cells can also produce a variety of control signals to regulate the proliferation and differentiation of the notch signaling pathway in central nervous system during development plays a vital role in the number of neuronal differentiation.This study, neural stem cell culture technology, the second cloning technology, and immunohistochemical techniques, RT-PCR technology to detect short-term removal of the EGF of FGF factor on neural stem cell proliferation and differentiation.Experimental procedure:sterile extract14days of gestation fetal rat cerebral cortical cells in primary suspension culture after3days, take part of the trypsin digestion as a single cell, the withdrawal of the factor of the EGF and FGF factor,1×105density seeded in96-well plates to continuesuspension culture for three days to observe the number of neurospheres cloning. Will train for three days neurospheres and digest the single cells of the neural stem cells were seeded in the coated with poly-ornithine+boric acid on the slide to make it adherent growth, and the withdrawal of the factor. Three days after the culture of pi/hochest staining compare the number of single-cell group of neural stem cells survive. Neurospheres cultured for three days and seven days a NF staining, and the neurospheres group of stem cell differentiation. And cultured neurospheres for three days and seven days the proportion of RT-PCR semi-quantitative detection of neurons and glial cell differentiation. The experimental results show that:isolated and cultured rat neural stem cells in suspension culture three days after the light microscope, the cells were round globular cell clusters, typical of neural stem cell characteristics and after nestin identification visible. By pi/hochest staining in the control group survival rate of92%, the withdrawal of the factor group survival rate was75%, the two groups there were significant differences (P<0.05) count results show that the withdrawal of the factor group of Neural Stem Cells in the number of clonedsignificantly less than the control group (P<0.05). Nf Dyeing, the withdrawal of neuronal differentiation factor group was significantly more in the control group, RT-PCR analysis results support the immunohistochemical staining results (P<0.05).With further research on the EGF and FGF in nerve cell differentiation and proliferation, looking forward to clear the mechanisms of neural stem cell proliferation and differentiation, thereby promoting the process of clinical application of stem cell transplantation for treatment of various neurodegen-erative diseases. The prospects for stem cell applications will be much broader.