Perfusion Culture Chip Providing Different Strengths of Shear Stress for Analysis of Vascular Endoth

• 作 者: Koji Hattori Hideki Kobayashi Shinji Sugiura Yoichi Munehira Toshiyuki Kanamori
• 机 构: National Institute of Advanced Industrial Science and Technology
• 出 处: 第八届全国微全分析系统学术会议、第三届全国微纳尺度生物分离分析学术会议暨第五届国际微化学与微系统学术会议
• 发表日期: 2013年2期
• 关键词: Perfusion culture Endothelial cell Microfluidic device Shear stress Quantitative

　　Endothelial cell (EC) in vivo is exposed to shear stress caused by blood flow and secretion activities of ECs are shear stress dependent [1].It is therefore important to culture ECs under the shear stress for drug discovery [2].In this paper,we developed a microfluidic device,which can control the shear stress in cell culture environment.The shear stress responsive genes were analyzed by PCR with the activated ECs after the perfusion culture.The perfusion culture chip was equipped with three perfusion culture units and HUVECs can be cultured by exposing three different strengths of shear stress (Fig.1).The microfluidic network of each perfusion culture unit is composed of ten "cell culture channels" (total area of culture surface: 12 cm2) and five "flow control channels".The flow control channels have three different depths.The perfusion rate of the medium in each cell culture channels is determined by the large fluidic resistances of the flow control channels and the pressure applied to the inlet ports.In the perfusion culture,the chip was placed in tight contact by a frame to prevent the deformation of the microchannels by a pressure.After the perfusion culture,HUVECs were harvested from the cell culture channels by the medium flow with trypsin/EDTA treatment and analyzed the gene expression by quantitative PCR.After the perfusion culture,HUVECs were aligned along the direction of medium flow (Fig.2).The mRNA expression levels of endothelial nitric oxide synthase (eNOS),thrombomodulin (THBD) in HUVECs were increased dependent of the strength of the applied shear stress (Fig.3).These results suggest that the shear stress was activated the vascular endothelial functions of HUVECs.The vascular endothelial functions of HUVECs activated by multi-shear conditions are successfully evaluated by not only cellular morphology but also PCR with the perfusion culture chip.The perfusion culture chip is expected to be applied to the screening for the vascular targeting drugs.