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Sensing system based on highly-functional cells on environmentally controlled microchips

基于环境控制微芯片上的高功能细胞的传感系统

基本信息

批准号：
13124204
负责人：
FUJITA Hiroyuki
金额：
$ 33.28万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13124204/
关键词：
MEMS fluidics nano biology Cell-based sensing cell culture surface modification

项目摘要

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.

该项目旨在在微加工芯片上培养高功能细胞，如肥大细胞和神经元细胞，并测量它们对化学刺激的反应。我们优化了表面改性。芯片结构和培养环境，以便在有限的空间中培养和维持健康细胞。(1)基于PDMS微流控通道的动物细胞过敏传感器研制了一种基于在PDMS微流控通道中培养的RBL-2 H3肥大细胞的过敏传感器。PDMS表面在氧等离子体下呈现亲水性，用于长期（超过一周）培养。成功地测量了使用奎纳克林荧光染料的肥大细胞的过敏反应，这证明了我们的过敏传感器的概念。(2)微阵列神经元芯片药物筛选反应器由1248个500 × 500 × 200立方微米的微腔组成。将来自鸡胚的原代培养神经元细胞与连接到每个室中的珠的各种肽x-x-x-x-x（x：Ile、Leu、Met、Phe、瓦尔）一起培养。某些类型的肽以与NGF（神经元生长因子）类似的方式诱导轴突生长。细胞外信号调节激酶也被激活。这表明使用细胞培养芯片进行药物筛选的可行性。(3)微通道中神经细胞的培养制作了正方形网格的微通道。将64个电极的阵列与放置和培养神经元细胞（原代培养细胞和PC 19细胞）的通道的交叉点对齐。它们沿着通道生长轴突，并根据通道连接形成网络。在四周内，从电极检测到一些神经元信号。(4)单个细胞处理芯片通过在芯片上制造的小孔抽吸，将许多细胞置于规则阵列中。通过与每个孔对齐的一对电极对每个细胞进行电穿孔。成功率为5- 10%。