title: [Study on in vitro isolation and culture method of neural stem cells from fetal rat neocortex].
abstract: OBJECTIVE
  To establish method for isolation and culture of neural stem cells from fetal rat neocortex in vitro.
  
  
  METHODS
  Neocortexes of fetal SD rats were isolated on gestational day 14. Mechanical triturations were used to improve cell dissociation status and conditions of serum-free media were optimized. Three test methods including neural stem cell marker protein (nestin and SOX2), proliferation and clonogenic ability and the multilineage differentiation potential were employed to identify neural stem cells.
  
  
  RESULTS
  Nestin of cells cultured with this method was expressed with strong positive, and more than 99% cells were SOX2-positive. BrdU was incorporated into the cells, and the amounts of the cells were 10.55 times as much as the initial cell number after 3 days of culture. The ratios of clonal neurosphere were (33.00 +/- 4.40)% after 6 days of culture. Staining with specific protein markers in the corresponding cells confirmed that neural stem cells could form neurons (MAP2), astrocytes (GFAP) and oligodendrocytes (O4) after differentiation induction.
  
  
  CONCLUSIONS
  Neural stem cells obtained with this method had high purity and cell output, potent self-renewal and multi-lineage differentiation ability, which will provide a good model for toxicological research.
authors.affiliation: Department of Environmental Health, School of Public Health, Fudan University, Key Laboratory of Public Health Safety, Ministry of Education, Shanghai 200032, China. xwang6@fudan.edu.cn
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.forename: Xiao
authors.initials: X
authors.lastname: Wei
authors.forename: Weiwei
authors.initials: W
authors.lastname: Zheng
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.forename: Hanyi
authors.initials: H
authors.lastname: Chen
authors.forename: Dajun
authors.initials: D
authors.lastname: Tian
authors.forename: Songhui
authors.initials: S
authors.lastname: Jiang
authors.forename: Weidong
authors.initials: W
authors.lastname: Qu
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005260:Female; D005333:Fetus; D007381:Intermediate Filament Proteins / Q000378:metabolism; D008297:Male; D019579:Neocortex / Q000166:cytology*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D058953:Neural Stem Cells / Q000166:cytology*; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D055748:SOXB1 Transcription Factors / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D055748:SOXB1 Transcription Factors; C527784:Sox2 protein, rat