Combination of basic fibroblast growth factor and brain-derived neurotrophic growth factor influences the differentiation of adult rat hippocampus neural stem cells into neurons
10.3969/j.issn.1673-8225.2009.27.027
- VernacularTitle:联合应用碱性成纤维细胞生长因子和脑源性神经营养因子对成年大鼠脑海马神经干细胞分化为神经元的影响
- Author:
Lei TONG
;
Dalong XIE
;
Hai GAO
;
Xiaojie TONG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(27):5331-5335
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Differentiation of neural stem calls (NSCs) was mediated by many environmental factors. Several factors can induce NSCs to differentiate into neurons in varying degrees and it is now a focus on the control of NSCs differentiation.OBJECTIVE: To study the effects of combination of basic fibroblast growth factor (bFGF) and brain-derived neurotrophic growth factor (BDNF) on the differentiation of NSCs into neurons.DESIGN, TIME AND SETTING: The in vitro cytology observation was performed at the Neurotomia Laboratory of China Medical University in May 2008.MATERIALS: Three adult male SD rats were provided by Experimental Animal Center of China Medical University.METHODS: Dispositions to the rats were consistent with ethical standards of animals. The rat brain hippocampus was removed sterilely. After trypsin digestion, NSCs were cultured in serum-free medium. Cell suspension was prepared and diluted when the diameter of the fourth passage of clone sphere was 200 μm by mixture of DMEM/F12 containing 2% B27, 20 μg/L of epidermal growth factor (EGF), and 20 μg/L bFGF. Monoclonal calls were passagad. NSCs were divided into blank control, bFGF, BDNF and bFGF+BDNF groups by different growth factors added into the media. Fetal bovine serum of 0.1 volume fraction was added in blank control group. The media in the other three groups were added bFGF, BDNF and bFGF+BDNF respectively for 1 week.The concentration of bFGF was 10 μg/L and the concentration of BDNF was 200 μg/L.MAIN OUTCOME MEASURES: Immunocytochemistry staining was used to identify NSCs as well as to detect the differentiation of NSCs into neurons.RESULTS: The monoclonal calls expressed nestin and the differentiated call expressed neuron specific enolase and glial fibrillary acidic protein. Compared to blank control group, the proportion NSCs into neurons in the bFGF group, BDNF group and bFGF+BDNF group were much higher (t=3.409-7.558, P < 0.05), with the highest in bFGF+BDNF group (t =7.558, P < 0.05).CONCLUSION: Combination of bFGF and BDNF can promote the differentiation of adult hippocampus NSCs into neurons.
