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Axonal Transport and Cytoskeleton

開催日 2014/9/13
時間 11:00 - 12:00
会場 Poster / Exhibition(Event Hall B)

SEM observation of neurons on various substrates and nano-structures

  • P3-017
  • 河西 奈保子 / Nahoko Kasai:1 後藤 東一郎 / Touichiro Goto:1 Lu Rick / Rick Lu:1 樫村 吉晃 / Yoshiaki Kashimura:1 田中 あや / Aya Tanaka:1 塚田 信吾 / Shingo Tsukada:1 住友 弘二 / Koji Sumitomo:1 
  • 1:NTT物性科学基礎研究所 / NTT Basic Res.Labs. 

To understand the mechanism behind the interaction between cells and substrates is important if we are to control neuronal growth and realize biological devices which function with living neurons. Our group has studied the chemical and physical conditions of substrates with the aim of examining and controlling axonal growth and synaptic formation.
In this study, we observed the surface morphology and cross-section of neurons cultivated on substrates made of different materials and with different structures using scanning electron microscopy (SEM). The cross-sections were obtained by using focused ion beam (FIB) milling technology.
Neurons were obtained from Wister rats and cultivated for 7 days on four different substrates; gold, titanium, indium tin oxide (ITO) pre-coated with cell adhesive matrix of laminin and poly-D-lysine, or a self-assembled monolayer (SAM) of hexanoic acid. Neurons were also cultivated on amorphous silicon or gold nano-pillars with different sizes, distances and heights. The neurons were chemically fixed and freeze dried using t-butanol.
The surface morphology obtained by SEM showed that the neurons cultivated on gold substrates were small and spherical. In contrast, the neurons cultivated on pre-coated ITO substrates were large and flatter. The cross-sectional observation demonstrated that the neurons cultivated on gold substrates were not completely attached to the substrate while the neurons cultivated on pre-coated ITO substrates adhered well and homogeneously to the substrate. The characteristics of the neurons cultivated on the titanium and SAM layer substrates fell between those of the neurons on the above two substrates. This insufficient attachment of neurons to the gold substrate could result in the construction of a complete cytoskeleton in the cell bodies, which would make the cell body small and spherical, and, furthermore, could remove cell bodies from the substrate. Neurons were successfully grown on a nano-pillar substrate, and showed better attachment to silicon nano-pillars than gold nano-pillars.
These results showed that neurons exhibit a clear preference of material, and imply the possibility of the neuronal control of neuronal growth by selecting proper materials and structures.

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