ISOLATION OF NEURAL STEM CELLS FROM NEONATAL RAT HIPPOCAMPUS AND THEIR IN VITRO DIFFERENTIATION INTO CHOLINERGIC NEURONS
- VernacularTitle:新生大鼠海马源性神经干细胞的分离培养及其向胆碱能神经元定向诱导分化研究
- Author:
Xiangying LUO
;
Zhimin YANG
;
Xiaobin SONG
;
Su LIU
;
Kuangyan ZHAO
;
Zhongtang FENG
;
Tinghua WANG
- Publication Type:Journal Article
- Keywords:
cell culture;
neural stem cell;
immunocytochemistry;
induction;
differentiation;
cholinergic neuron
- From:
Chinese Journal of Neuroanatomy
2005;21(2):190-194
- CountryChina
- Language:Chinese
-
Abstract:
The present study aims to isolate neural stem cells from neonatal rat hippocampus and induce them to differentiate into cholinergic neurons. A multipotent cell line derived from the hippocampi of neonatal rats which had the ability to form clones was incubated in serum-free DMEM/F12 medium added with 20ng/ml basic fibroblast growth factor (bFGF) and B27. After differentiation of the neural stem cells, immunocytochemistry was used to detect nestin, the antigen of the cell clone, and β-tubulin (Tuj 1 ), glial fibrillary acidic protein (GFAP) and galactocerebroside (Galc), the markers specific for neurons, astrocytes and oligodendrocytes, respectively. Embryonic chick skeletal muscle extract was used to induce the differentiation of the neural stem cells into cholinergic neurons. The results showed that the cell line isolated from the hippocampi of neonatal rats expressed nestin and had the potential to form clones and differentiate into neurons, astrocytes and oligodendrocytes. Embryonic chick skeletal muscle extract can induce 9.6% of the isolated cell line to differentiate into cholinergic neurons compared with 3.9% in controls. These findings suggested that the cell line, which expressed nestin antigen, was a multipotent cell line capable of self-renewing, and was believed to contain stem cells of the CNS. These neural stem cells can be induced to differentiate into cholinergic neurons by using embryonic chick skeletal muscle extract.
