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2022, Vol. 26 ›› Issue (13): 2069-2074Expression and function of colony-stimulating factor 1 in differentiation of neural stem cells induced by activated astrocyte conditioned mediumShan Wen, Shi Wei
Abstract: BACKGROUND: The excessive proliferation of reactive astrocytes after traumatic brain injury may have an impact on the differentiation of neural stem cells into neurons. Therefore, studying the regulation of reactive astrocytes on neural stem cell differentiation after traumatic brain injury and its regulation mechanism may have great clinical application prospect for promoting the recovery of neural function. OBJECTIVE: To investigate the effect of reactive astrocytes on the differentiation of neural stem cells into neurons. METHODS: Astrocytes of SD rats were cultured in vitro and divided into control group and lipopolysaccharide stimulation group. After 3 days of intervention, the expression of colony-stimulating factor 1 in astrocytes was detected by PCR, ELISA and western blot assay. Primary neural stem cells of SD rats were cultured in vitro, and astrocyte conditioned medium was added to the neuron induction medium for 3 days. Immunofluorescence and flow cytometry were used to detect the expression of β-tubulin III and glial fibrillary acidic protein. Primary neural stem cells were cultured in vitro, and the recombinant protein of colony-stimulating factor-1 was added to the neuron induction medium for 3 days. The effect of colony-stimulating factor-1 on the differentiation of neural stem cells into neurons was detected by flow cytometry and immunofluorescence. RESULTS AND CONCLUSION: (1) The expression of colony-stimulating factor-1 in astrocytes stimulated by lipopolysaccharide and in conditioned medium was very low. (2) The coculture of neural stem cells and astrocyte conditioned medium stimulated by lipopolysaccharide inhibited the differentiation of neural stem cells into neurons. (3) Neural stem cells promoted the differentiation of neural stem cells into neurons by adding colony stimulating factor 1 recombinant protein. (4) The results showed that colony-stimulating factor 1 secretion of astrocytes was down-regulated after stimulation with lipopolysaccharide, and inhibited the differentiation of neural stem cells into neurons. The overexpression of colony-stimulating factor 1 can promote the differentiation of neural stem cells into neurons, which can repair brain injury in rats. Key words:stem cells, neural stem cell, traumatic brain injury, lipopolysaccharide, astrocytes, colony-stimulating factor 1, nerve repair |