Numerical analysis of on-chip microdroplet electroporation process and clarification of the mechanism

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K04183
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 19010:Fluid engineering-related
• Research Institution: Sasebo National College of Technology
• Principal Investigator: Nakashima Kenji 佐世保工業高等専門学校, 機械工学科, 教授 (40311112)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 数値解析 / 液滴エレクトロポレーション / マイクロ流体デバイス / 細胞膜穿孔 / 遺伝子導入 / 液滴形成 / ディーン流れ / 電気穿孔

Outline of Final Research Achievements

To develop an "on-chip microdroplet electroporation process" that can efficiently establish iPS cells, this study visualizes the phenomenon using numerical analysis. Therefore, the numerical analysis was divided into four stages: (1) cell alignment by Dean flow, (2) droplet formation to separate cells one by one, (3) membrane electron permeabilization in electroporation, and (4) gene transfer by electrophoresis. (1) It was confirmed that the velocity of droplets passing over the electrode was not sufficient to achieve the effect of cell alignment by Dean flow. (2) Characteristic distributions of interfacial tension and viscosity were obtained to enable stable droplet formation. (3) The characteristics of pore formation with changes in the position of cells in the droplet were organized. (4) The calculation could be performed using a simple method based on the calculation by mobility.

Free Research Field

流体

Academic Significance and Societal Importance of the Research Achievements

本研究で扱う「オンチップ微小液滴電気穿孔法」とは，細胞への遺伝子導入操作（電気穿孔）を連続的に施す技術である．この工程は，幅50μm×高さ30μmの矩形流路で構成される，全長2cm以内に収まるマイクロ流体デバイスによって実現される．この技術によって，安定的にiPS細胞の樹立が可能になれば，現在主流のウィルスベクター法による技術に置き換えることが可能になる．そして，本研究で開発する手法は，大規模並列処理と相性が良い．それゆえ，産業的にiPS細胞を大量生産することが可能になるため，iPS細胞を用いた臨床応用や製薬開発の推進に貢献することが可能となる．