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.

该项目旨在在微机械芯片上培养高功能细胞，例如肥大细胞和神经元细胞，并测量其针对化学刺激的反应。我们已经优化了表面修饰。芯片结构和文化环境为了在狭窄的空间中培养和维持健康细胞。（1）在PDMS微流体通道中使用动物细胞的过敏传感器基于在PDMS（POLYDIMETHYLSILOXANE）微流体通道中培养的RBL-2H3肥大细胞的过敏传感器。将PDMS表面在氧血浆中呈现至长期（一周）培养物的亲水性。使用喹分河荧光染料对肥大细胞的反应进行了成功测量。这证明了我们对过敏传感器的概念。（2）用微阵列神经元芯片进行药物筛选。阵列反应器由1248个微室组成，大小为500x500x200立方微米。用各种肽X-X-X-X-X（X：Ile，Leu，Met，Phe，Phe，Val）培养来自鸡胚胎的原发性培养神经元细胞。某些类型的肽以与NGF相似的方式诱导轴突生长（神经元生长因子）。还激活了细胞外信号调节激酶。这表明使用细胞培养芯片进行药物筛查的可行性。（3）微通道中神经元细胞的培养物制造了微通道的平方晶格。将64个电极的阵列与位于神经元细胞（原代培养细胞和PC19细胞）的通道的交点对齐并进行培养。它们根据通道连接沿通道沿通道并形成网络。在四个星期的时间内从电极中检测到一些神经元信号。（4）单个细胞交接芯片通过在芯片中制成的小孔中抽吸将许多细胞放在常规阵列中。对每个电池的电穿孔是通过对齐每个孔对齐的一对电极进行的。获得5-10％的成功率。