title: Culture and differentiation of rat neural stem/progenitor cells in a three-dimensional collagen scaffold.
abstract: A stable and fast method for constructing a neural-like tissue from rat neural stem/progenitor cells (rNS/PCs) based on three-dimensional (3D) collagen gel is described. First step, the collagen-embedded rNS/PCs expanded with the medium consisting of DMEM/F12/RPMI1640 (1:1:1) supplemented with EGF and bFGF was used to expand the cells in gel in 96-well plates until the average diameter of cell clusters was about 50-100 μm with the cell density higher than 10(7) cells/mL. In the second step, the initial medium was replaced with NB/B-27 supplemented with bFGF and BDNF. The results show that cells in collagen presented neural-like morphology and maintained live cell rate around 82 % in neural network pattern at least for 42 days under static conditions. The cell-collagen constructs were detected by immunofluorescence and immunohistochemistry test after 42 days of culture, part of cells still maintained the character of rNS/PCs, and others differentiated into neurons, astrocytes, and oligodendrocytes. Our 3D neural-like tissue construct was similar to the neural tissue in morphology and cell compositions. They thus have a potential to be used for drug screening, detection of environment toxins, and replacement therapy.
authors.affiliation: Dalian R & D Center for Stem Cell and Tissue Engineering, Chemical Engineering School, Dalian University of Technology, Dalian, 116024, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Ge
authors.forename: Kedong
authors.initials: K
authors.lastname: Song
authors.forename: Shui
authors.initials: S
authors.lastname: Guan
authors.forename: Yanli
authors.initials: Y
authors.lastname: Qi
authors.forename: Bo
authors.initials: B
authors.lastname: Guan
authors.forename: Wenfang
authors.initials: W
authors.lastname: Li
authors.forename: Junshan
authors.initials: J
authors.lastname: Liu
authors.forename: Xuehu
authors.initials: X
authors.lastname: Ma
authors.forename: Tianqing
authors.initials: T
authors.lastname: Liu
authors.forename: Zhanfeng
authors.initials: Z
authors.lastname: Cui
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D003470:Culture Media / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D054457:Tissue Scaffolds
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D003470:Culture Media; D016222:Fibroblast Growth Factor 2; D003094:Collagen