10:00 〜 10:15
▲ [14a-3A-4] Neuronal selective growth on nanopillars using supported lipid bilayer
キーワード：nanopillar,neuron,supported lipid bilayer
Neuronal patterning is of interest for understanding signal propagation between neurons and neuron-matrix interactions and thus elucidating neuronal functions. It is also useful for cell-based assays. The patterning of living cells in vitro has been made possible thanks to surface topographic features and/or physicochemical properties. In this study, the neurons were successfully patterned on nanometer-scale pillars used as scaffolds using a supported lipid bilayer (SLB) covering a base substrate to avoid cellular attachment.
500nm-diameter nanopillars of amorphous silicon (a-Si) were fabricated on a quartz substrate using electron beam lithography. For neuronal growth selectivity towards the pillar, an SLB composed of 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) was formed using lipid self-spreading to avoid cell attachment. Neurons were prepared from rat cortex, and cultivated on the above nanopillar substrate with the SLB for 7-9 days. Neuronal growth was examined by fluorescent immunostaining using confocal microscopy and scanning electron microscopy (SEM).
We confirmed using a fluorescent microscope that the SLB was formed by self-spreading. Then SEM observation revealed that the neurons of 7DIV grew selectively on the nanopillars. However, the neurons grew randomly where no SLB was formed.
These results show that the non-fouling characteristic of an SLB is useful for patterning neurons on nanopillars used as scaffolds and for providing an in vitro biological environment in which to examine neuronal functions.
500nm-diameter nanopillars of amorphous silicon (a-Si) were fabricated on a quartz substrate using electron beam lithography. For neuronal growth selectivity towards the pillar, an SLB composed of 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) was formed using lipid self-spreading to avoid cell attachment. Neurons were prepared from rat cortex, and cultivated on the above nanopillar substrate with the SLB for 7-9 days. Neuronal growth was examined by fluorescent immunostaining using confocal microscopy and scanning electron microscopy (SEM).
We confirmed using a fluorescent microscope that the SLB was formed by self-spreading. Then SEM observation revealed that the neurons of 7DIV grew selectively on the nanopillars. However, the neurons grew randomly where no SLB was formed.
These results show that the non-fouling characteristic of an SLB is useful for patterning neurons on nanopillars used as scaffolds and for providing an in vitro biological environment in which to examine neuronal functions.