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

该项目的目标是在微机械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。我们对表面改性进行了优化。芯片结构和培养环境，以便在有限的空间内培养和维持健康细胞。(1)利用PDMS微流控通道中的动物细胞研制了基于PDMS(聚二甲基硅氧烷)微流控通道中培养的RBL-2H3肥大细胞的变态反应传感器。PDMS表面在氧气等离子体下呈现亲水性，进行长期(超过一周)的培养。利用喹啉荧光染料成功地测量了肥大细胞的反应，验证了我们提出的过敏传感器的概念。(2)微阵列神经元芯片药物筛选阵列反应器由1248个500×500×200立方微米的微室组成。原代培养的鸡胚神经细胞在每个小室中分别加入不同多肽x-x(x：Ile、Leu、Met、Phe、Val)进行培养。某些类型的多肽以类似于神经生长因子(NGF)的方式诱导轴突生长。细胞外信号调节激酶也被激活。这表明利用细胞培养芯片进行药物筛选是可行的。(3)神经细胞在微通道中的培养制备了方形晶格的微通道。将电极阵列与放置和培养神经元细胞(原代培养细胞和PC19细胞)的通道相交。它们沿着通道生长轴突，并根据通道连接形成网络。4.单个细胞处理芯片将许多细胞通过芯片上的小孔抽吸成规则的阵列。通过与每个孔对齐的一对电极对每个电池进行电穿孔。成功率为5-10%。

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.