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Culture and Differentiation of Rat Neural Stem/Progenitor Cells in a Three-Dimensional Collagen Scaffold

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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 107 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.

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Acknowledgment

This work was supported by Fok Ying Tung Education Foundation (132027), National Science Foundation of China (31170945), Dalian Science and Technology Plan (Biology and Medicine, 2012E15SF174) and Fundamental Research Funds for the Central Universities (DUT11SM06/09/12JB02/09) and Project-sponsored by SRF for ROCS, SEM and the State Key Laboratory of Fine Chemicals (KF1111).

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Authors and Affiliations

  1. Dalian R & D Center for Stem Cell and Tissue Engineering, Chemical Engineering School, Dalian University of Technology, Dalian, 116024, China

    Dan Ge, Kedong Song, Shui Guan, Yanli Qi, Bo Guan, Wenfang Li, Xuehu Ma & Tianqing Liu

  2. Key Laboratory for Micro/Nano Technology and System of Liaoning Province, Dalian University of Technology, Dalian, 116024, China

    Junshan Liu

  3. Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, Oxford, UK

    Zhanfeng Cui

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  1. Dan Ge
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  2. Kedong Song
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  3. Shui Guan
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  4. Yanli Qi
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  5. Bo Guan
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  6. Wenfang Li
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  7. Junshan Liu
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  8. Xuehu Ma
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  9. Tianqing Liu
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  10. Zhanfeng Cui
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Correspondence to Tianqing Liu.

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Ge, D., Song, K., Guan, S. et al. Culture and Differentiation of Rat Neural Stem/Progenitor Cells in a Three-Dimensional Collagen Scaffold. Appl Biochem Biotechnol 170, 406–419 (2013). https://doi.org/10.1007/s12010-013-0211-5

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  • DOI: https://doi.org/10.1007/s12010-013-0211-5

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