| Research Abstract |
This project aimed to culture highly-functional cells, such as mast cells and neuron cells, on micromachined chips and to measure their response against chemical stimulus. We have optimized the surface modification. chip structures and cultural environment in order to culture and sustain healthy cells in a confined space.
(1) Allergy sensor using animal cells in PDMS microfluidic channel An allergy sensor based on RBL-2H3 mast cells cultured in a PDMS (polydimethylsiloxane) microfluidic channel was developed. The PDMS surface was rendered to hydrophilic under oxygen plasma for long-term (over a week) culture. Alergic, response of mast cells using quinacrine fluorescent dye was successfully measured ; this proved our concept of the allergy sensor.
(2) Drug screening with microarrayed neuron chip The arrayed reactor was composed of 1248 micro chambers of 500x500x200 cubic micrometers in size. Primary culture neuron cells from a chicken embryo were cultured with variety of peptides x-x-x-x-x (x : Ile, Leu, Met, Phe, Val) attached to a bead in each chamber. Certain types of peptides induced axon growth in the similar manner to the NGF (neuron growth factor). Extracellular signal-regulated kinase was also activated. This suggests the feasibility of using a cell culture chip for drug screening.
(3) Culture of neuron cells in micro channels A square lattice of microchannels were fabricated. An array of 64 electrodes was aligned with the intersection of channels where neuron cells (primary culture cells and PC19 cells) were placed and cultured. They grew axons along the channel and formed networks according to the channel connection. Some neuron signals were detected from electrodes over four weeks.
(4) Individual cell handing chip Many cells were placed in a regular array by aspiration through small holes made in a chip. Electroporation to each cell was conducted by a pair of electrodes aligned to each hole. Success rate of 5-10 % was obtained.
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