Abstract:

BACKGROUND: Directed differentiation and proliferation of nerve stem cells are regulated by their own genes and external signals.
OBJECTIVE: To observe the differentiation of mesencephalic neural stem cells into dopaminergic neurons under normoxic and hypoxic conditions induced by glial-derived neurotrophic factor (GDNF).
METHODS: Under aseptic conditions, we isolated mesencephalic tissues from E12 mouse embryos. Single cell suspension was prepared by pancreas enzyme digestion and mechanical pipetting, and were cultured and proliferated in serum-free medium. Nestin immunocytochemistry staining was used to identify nerve stem cells. Purified nerve stem cells differentiated in serum medium. Neuron-specific enolase and glial fibrillary acidic protein immunocytochemistry staining methods were performed to identify neurons and astrocytes, respectively. All the cells were divided into normoxia group, normoxia+GDNF group, hypoxia group, hypoxia+GDNF group, and then cultured in serum medium.
RESULTS AND CONCLUSION: Under the hypoxia condition, mesencephalic neural stem cells had a higher differentiated rate than that in the normoxia group. The number of differentiated cells in the hypoxia+GDNF group was higher than that of the other groups, and the differentiated cells had a mature phenotype. It is indicated that under the hypoxic condition, GDNF can extremely promote the differentiation of nerve stem cells into dopaminergic neurons with good morphology and mature function.