Astrocyte-derived extracellular vesicles promote the differentiation of rat neural stem cells into neurons

doi:10.12307/2023.672

Abstract

Abstract: BACKGROUND: Neural stem cells have the potential to differentiate into neurons. It has been reported that astrocytes are closely related to neurons, but whether astrocyte-derived extracellular vesicles can promote the efficient differentiation of neural stem cells into neurons has not been reported.
OBJECTIVE: To explore the possibility of inducing and promoting neuronal differentiation of neural stem cells from extracellular vesicles derived from astrocytes.
METHODS: (1) Rat neural stem cells and astrocytes were cultured in vitro and identified by cell morphology and immunofluorescence. Astrocyte-derived extracellular vesicles were further extracted and identified by transmission electron microscopy, particle size analysis, western blot assay and other methods. (2) Passage 3 neural stem cells were divided into two groups: Neural stem cells were treated with the same volume of PBS and astrocyte-derived extracellular vesicles (1 mg/mL) for 6 days. (3) The expression levels of neuron markers after astrocyte-derived extracellular vesicle intervention were detected by immunofluorescence, qRT-PCR and western blot assay.
RESULTS AND CONCLUSION: (1) Under the microscope, neural stem cells were spherical in shape and distributed in cell clusters. Immunofluorescence showed that the characteristic markers CD133 and Nestin were highly expressed. (2) Astrocyte morphology was irregular stellate under the microscope, and immunofluorescence showed that the characteristic marker glial fibrillary acidic protein was highly expressed. (3) The shape of astrocyte-derived extracellular vesicles was round or oval cup and 10-200 nm in diameter by dynamic light particle scatterer. Western blot assay showed positive expression of CD63 and TSG101, related markers of extracellular vesicles. (4) Compared with PBS group, astrocyte-derived extracellular vesicle group showed a large number of adherent differentiation of neural stem cells, and the differentiated cells were plump and obvious protrusions, which were mainly neuron-like, and the expression of tubulin β3 and neurofilament 200 was high (P < 0.05). (5) These results suggest that astrocyte-derived extracellular vesicles can promote the differentiation of neural stem cells into neurons.

Key words: neural stem cell, astrocyte, extracellular vesicle, central nervous system disease, neuron, differentiation, spinal cord injury, nerve repair

CLC Number:

Cai Chuang, Huang Yonghui, Cao Xingbing, Sun Haitao, Guan Shihao, Huang Wenkang, Han Bo. Astrocyte-derived extracellular vesicles promote the differentiation of rat neural stem cells into neurons[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(24): 3845-3851.

Figures/Tables 6

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