Effects of folic acid on in vitro astrocytic differentiation of neural stem cells from neonatal rats  

作　　者：Xumei Zhang Guowei Huang Zhihong Tian Guanglei Wang Dalin Ren 

机构地区：[1]Department of Nutrition and Food Hygiene, School of Pubfic Health, Tianjin Medical University, Tianjin 300070, China

出　　处：《Neural Regeneration Research》2009年第8期613-617,共5页中国神经再生研究（英文版）

基　　金：the National Natural Science Foundation of China,No.30571563, 30771797;the grant from Tianjin Education Commission,China,No. 20070208

摘　　要：BACKGROUND： Folic acid is essential for normal functioning of the nervous system. Previous studies have focused on the effects of folic acid on astrocyte proliferation. OBJECTIVE： To explore the effects of folic acid on astrocyte differentiation of neural stem cells （NSCs） and the related mechanisms in vitro. DESIGN, TIME AND SETTING： A randomized, controlled, grouping experiment was performed in Tianjin Medical University between August 2007 and October 2008. MATERIALS： Folic acid and 5-bromo-2-deoxyuridine （BrdU） were obtained from Sigma, MO, USA. Primary antibodies [rabbit anti-rat nestin, β-tubulin-Ⅲ, glial fibrillary acidic protein, and neurogeninl （Ngnl）; mouse anti-rat BrdU and β-actin monoclonal antibodies] were purchased from Santa Cruz Biotechnology, USA. METHODS： At 6 days of NSC proliferation from 24-hour-old neonatal rats, BrdU incorporation assay was performed. Seven days after primary culture, NSCs were induced to differentiate with medium containing 5% fetal bovine serum. Cultured NSCs were assigned to three groups： control, low-dose （liquid media with 8 mg/L folic acid）, and high-dose folic acid （liquid media with 44 mg/L folic acid）. MAIN OUTCOME MEASURES： At day 7 after primary culture, the cells were identified as NSCs by immunocytochemical methods. Double-label immunofluorescence technique for glial fibrillary acidic protein/BrdU detected differentiated cells 7 days after induction. Western blot was used to analyze expression of Ngnl protein in NSCs. RESULTS： In serum-free suspension medium, neurospheres comprised a large number of Nestin-, glial fibrillary acidic protein-, β-tubulin-Ⅲ-, and BrdU-positive cells. Compared with the control group, high-dose folic acid supplementation led to an marked increase in the percentage of glial fibrillary acidic protein/BrdU-positive cells （P 〈 0.05）, and significantly decreased Ngnl protein expression （P 〈 0.05）. CONCLUSION： Folic acid promotes astrocytic differentiation of NSCs, which might be reBACKGROUND： Folic acid is essential for normal functioning of the nervous system. Previous studies have focused on the effects of folic acid on astrocyte proliferation. OBJECTIVE： To explore the effects of folic acid on astrocyte differentiation of neural stem cells （NSCs） and the related mechanisms in vitro. DESIGN, TIME AND SETTING： A randomized, controlled, grouping experiment was performed in Tianjin Medical University between August 2007 and October 2008. MATERIALS： Folic acid and 5-bromo-2-deoxyuridine （BrdU） were obtained from Sigma, MO, USA. Primary antibodies [rabbit anti-rat nestin, β-tubulin-Ⅲ, glial fibrillary acidic protein, and neurogeninl （Ngnl）; mouse anti-rat BrdU and β-actin monoclonal antibodies] were purchased from Santa Cruz Biotechnology, USA. METHODS： At 6 days of NSC proliferation from 24-hour-old neonatal rats, BrdU incorporation assay was performed. Seven days after primary culture, NSCs were induced to differentiate with medium containing 5% fetal bovine serum. Cultured NSCs were assigned to three groups： control, low-dose （liquid media with 8 mg/L folic acid）, and high-dose folic acid （liquid media with 44 mg/L folic acid）. MAIN OUTCOME MEASURES： At day 7 after primary culture, the cells were identified as NSCs by immunocytochemical methods. Double-label immunofluorescence technique for glial fibrillary acidic protein/BrdU detected differentiated cells 7 days after induction. Western blot was used to analyze expression of Ngnl protein in NSCs. RESULTS： In serum-free suspension medium, neurospheres comprised a large number of Nestin-, glial fibrillary acidic protein-, β-tubulin-Ⅲ-, and BrdU-positive cells. Compared with the control group, high-dose folic acid supplementation led to an marked increase in the percentage of glial fibrillary acidic protein/BrdU-positive cells （P 〈 0.05）, and significantly decreased Ngnl protein expression （P 〈 0.05）. CONCLUSION： Folic acid promotes astrocytic differentiation of NSCs, which might be re

关 键 词：folic acid neural stem cells ASTROCYTE neurogenin 1 in vitro 

分 类 号：R741[医药卫生—神经病学与精神病学]