Cell array preparation in flow-type microchip by using photoresponsive polymer substrate
Integration of life sciences & engineering
Bio-transformation in the Laboratory and in Large Scale Production (T5-3)
Keywords: cell array, microchip, photoresponsive culture substrate, PEG, animal cell
Jun-ichi Edahiro†, Kimio Sumaru*†, Yuichi Tada†‡, Kyoko Kikuchi†, Yuki Oshima†, Shinji Sugiura†, Toshiyuki Takagi†, Toshio Shinbo†, Yasuo Yoshimi‡, Toshiyuki Kanamori†
† Research center of advanced bionics, national institute of advanced industrial science and technology (AIST), Central 5th, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, JAPAN
‡ Shibaura Institute of Technology, 3-9-14 Shibaura Minato-ku, Tokyo 108-8548, JAPAN
*k.sumaru@aist.go.jp
In recent years, cell array chips have been studied actively aiming at effective analysis of pharmacologic effect of bioactive substances. In order to make such analysis more efficient, we examined a novel layout of micro cell chip, in which living cells were arranged in the prescribed locations in microchannels. For this purpose, we developed a functional culture substrate on which cell adhering region can be prepared by regional light irradiation. The photoresponsive culture substrate was prepared on basal polystyrene plate by coating a photochromic polymer, polymethylmethacrylate modified with 6-nitrospiropyran, and polyethyleneglycol (PEG), which has a strong hindering effect on cell adhesion. When ultraviolet light (UV) with the wavelength of 365 nm was irradiated on the photoresponsive culture substrate in water, PEG was lost through the dissolution into water due to change in the interaction with the photochromic polymer. In this condition, adhering animal cells can be attached only at the UV-irradiated region.
In this study, we put a piece of polydimethylsiloxane (PDMS) having microchannel structure (width 600 μm, depth 200 μm) onto this photoresponsive culture substrate, and constructed a channel-type microchip. Since PDMS is transparent for UV irradiation, cell adhering region can be prepared by light irradiation even in the closed microchannel. Additionally, PDMS microchip is also feasible to culture cells for a long time since oxygen can be supplied through the PDMS by diffusion. In this microchannel, we confirmed that cell colonies of different animal cell lines; CHO-K1 or MDCK cells, can be arranged separately by using the local light irradiation. We expect that this technique can be used as a preparation method of cell array chip.
Presented Thursday 20, 15:40 to 16:00, in session Bio-transformation in the Laboratory and in Large Scale Production (T5-3).
