Yuki SHIRAISHI, Hiroaki SHINOHARA, and Minoru SUGA
Graduate School of Innovative Life Science, University of Toyama, 930-8555, Toyama City Japan.
Abstract: In this paper, we applied a two dimensional surface plasmon resonance (2D-SPR) system for insulin sensing by label-free observation of individual rat L6 myoblast cells (L6 cell) upon insulin stimulation. The L6 cells were cultured on a gold thin layer-coated glass chip. Individual L6 cells on the sensor chip were observed with a 2D-SPR sensor upon insulin stimulation. Insulin was detected in the concentration range from 5 to 100 μM by reflection intensity change after 50 min from insulin injection. We further considered the SPR response mechanism for insulin. The SPR signal response for insulin decreased in the presence of staurosporine or cytochalasin B. From these inhibitor experiments, we considered SPR response might reflect the insulin-induced initial kinase aggregation and following intracellular reactions such as actin-movement near the cell-bottom membrane. This L6 cell-based 2D-SPR system showed the usefulness of SPR observation for the label-free, simple, rapid, biosensing to various hormones with mammalian cells as sensing element and for the consideration of the intracellular reaction mechanism.
Keywords: 2D-SPR, Insulin sensing, Rat myoblast, Intracellular reactions, Label-free observation techniques
Pages: 23 – 31 | Full PDF Paper