Dissertation > Medicine, health > Basic Medical > Human morphology > Human histology

Population Expansion of Rat Hippocampal Neural Stem Cells and Their Integration with Three-dimensional Microwell Patterns

Author LvYanLing
Tutor WuZeZhi
School Chongqing University
Course Cell Biology
Keywords Hippocampal neural stem cells Neurospheres Amplification Polylactic acid Three - dimensional microstructures
CLC R329
Type Master's thesis
Year 2010
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Neural stem cells (neural stem cells, NSCs) are pluripotent stem cells derived from the central nervous system and play an important role in neural development and injury repair in stem cell research lies to obtain effective amplification methods have been created. Neural stem cell amplification including amplified under normal culture conditions, and under certain conditions in a three-dimensional scaffold amplification. Amplification of the reported neural stem cells in a three-dimensional scaffold, morphology and ion channel functional expression of behavioral characteristics different from the ordinary two-dimensional culture conditions, more close to the features under in vivo conditions. Therefore, the research of effective extraction methods and three-dimensional micro-structure of PLLA composite hippocampal stem cells. Objective To compare the growth characteristics of the two kinds of medium hippocampal neural stem cells (neural stem cells, NSCs), hippocampus neural stem cells (NSCs) optimization of the amplification and its three-dimensional polylactic acid (PLLA) the slight concave schema composite initial screening favorable for neural stem cells combined with stem cell characteristics to maintain the three-dimensional micro-structural configuration. Isolated rat embryonic hippocampal cells and using Neurobasal based medium, DMEM? F12 based medium for amplification. Methyl thiazolyl tetrazolium blue (MTT) assay and the number of neurospheres statistical method cell proliferation behavior of two kinds of medium. Ultraviolet lithography, silicon etching and soft lithography technology to prepare PLLA 3D microscopic concave Schema, and hippocampal NSCs with tiny concave schema composite. Using an optical microscope to detect the behavior of cell growth in the tiny concave Figure ceremony, and immunofluorescence identified differentiation in different three-dimensional structure. Results the primary separation hippocampal cells were neural stem cell marker nestin-positive to the differentiation the Department of neurons and glial cells. Amplification time of 30 d, hippocampal neural stem cells in Neurobasal-based medium slow amplification aggregate into neurospheres rare cell adherence and differentiation; to DMEM? F12 based medium hippocampal neural stem cells rapid amplification, but easy-adherent and differentiation. The number of neurospheres cultured day 25 the latter for the former and 4.7-fold. Therefore, the sequential application of two media can achieve effective amplification of hippocampal stem cells. Slight concave micromachining prepared schematic structure is clear and stable, with the structure of the high aspect ratio (≥ 1). Amplification of hippocampal neural stem cells in three-dimensional the tiny concave Figure ceremony successful composite growth of micro-structure the slight concave diameter as well as the presence and width of the channel significantly affect the number of cell growth in the micro-structure. With the increase of the diameter of the fine recessed corresponding increase in the number of growing cells in the micro-structure; With the introduction of the passage structure between the fine concave and the channel width is increased from 20μm to 40μm, the number of cells is also increased correspondingly. Cells in the micro-structure of the non-channel basic the neurites extended; lend a lot of nerve channel micro-structure of cell protrusions and projections extending along the microchannel. Conclusion DMEM? F12-based medium conducive to rat hippocampal NSCs amplification, Neurobasal-based medium conducive to the purification of the hippocampal NSCs. Sequential application of two kinds of medium hippocampal neural stem cells with three-dimensional composite the tiny concave Figure-effective amplification. No channel micro structure conducive to retaining characteristics of stem cells, the micro-structure of the channel to facilitate the differentiation of stem cells and neurites extending along the channel direction.

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