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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.

Original publication

DOI

10.1007/s12010-013-0211-5

Type

Journal article

Journal

Appl Biochem Biotechnol

Publication Date

05/2013

Volume

170

Pages

406 - 419

Keywords

Animals, Astrocytes, Brain-Derived Neurotrophic Factor, Cell Count, Cell Culture Techniques, Cell Differentiation, Cell Proliferation, Cell Shape, Cell Survival, Cells, Cultured, Collagen, Culture Media, Fibroblast Growth Factor 2, Hippocampus, Imaging, Three-Dimensional, Neural Stem Cells, Neurons, Rats, Rats, Sprague-Dawley, Tissue Scaffolds