Development of a multi-point sampling device for secreted cellular molecules to analyze intercellular communication

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K15426
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 28050:Nano/micro-systems-related
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Ota Nobutoshi 国立研究開発法人理化学研究所, 生命機能科学研究センター, 研究員 (60705036)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: マイクロバンドルプローブ / キャピラリー / 微量液体採取 / 細胞分泌物 / 局所刺激

Outline of Final Research Achievements

This study has aimed to develop a capillary-based open microfluidic device that can collect tiny amount of liquid from multiple regions with arbitrary number of sampling times, which promotes simple sampling and analysis of secreted molecules from multiple cell groups. By arbitrarily setting the positions of the capillary tips as well as assigning inlets and outlets, this device can determine the area of cells stimulated by introduced liquid and the range of liquid collected from vicinity of the cells. Cell removal through this device was performed by applying trypsin solution to confluent myoblasts on a culture dish. Minimum area of local myoblast removal was 0.004 square mm. In addition, the device showed its capability to make two adjacent groups of cultured macrophages that were stimulated or unstimulated by lipopolysaccharide, resulting in significantly different amount of secretion shown in time-course measurements.

Free Research Field

ナノマイクロシステム

Academic Significance and Societal Importance of the Research Achievements

細胞分泌物の採取は手作業で行われることも多く、一様に培養された細胞の中から特定の細胞の分泌物を狙って採取することは難しい。そのため通例、採取される細胞分泌物は培養容器中の全細胞に由来するものである。本研究のデバイスは、特定の細胞に由来する細胞分泌物を複数回にわたって採取することを可能とするため、細胞間で行われる細胞分泌物によるコミュニケーションの解析に使用できる。さらに、マイクロ流体制御を通じて特定の細胞に刺激を与えられるため、細胞パターニングなどの細胞組織工学や、例えば初期がん細胞の発生のような細胞組織の初期発生研究に応用できる。