2022 Fiscal Year Final Research Report
Effective fabrication and selective retrieval of hybridomas from among cell populations
| Project/Area Number |
20H02771
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Review Section |
Basic Section 34020:Analytical chemistry-related
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| Research Institution | University of Hyogo |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
鈴木 雅登 兵庫県立大学, 理学研究科, 准教授 (60574796)
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Keywords | 誘電泳動 / 細胞アレイ / ハイブリドーマ / 細胞融合 / 選択回収 |
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| Outline of Final Research Achievements |
Dielectrophoresis (DEP) was used to trap the target B cells from the splenocyte populations in microwell arrays to select useful cells. Here, microparticles were attached to the cells except target B cells to change their dielectrophoretic behavior. The useful B cells with antibody-secreting ability trapped in the microwells were discriminated. Antigens immobilized on the bottom of wells or on the cell surface were used to capture the antibodies secreted from the cells. Labeling of the captured antibodies by a fluorescence molecule allowed us to evaluate the antibody-secreting ability of individual cells easily. After both B cells and myeloma cells were trapped in the wells, asymmetric electric field pulses were applied to form heterogeneous cell fusions of different diameters with suppressed cell disruption. B cells with antibody-secreting ability were selectively retrieved by using negative DEP to the wells.
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| Free Research Field |
バイオ分析化学
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では，大量数のリンパ球の中から，標的とする細胞のみを捕捉，捕捉細胞の機能評価，細胞融合，標的細胞の回収を，迅速で簡便に達成できるデバイスの開発を行った．これまでのウェルアレイを用いた細胞選択では，数十万個のウェルに候補の細胞を捕捉し，わずかに含まれる抗体産生細胞を蛍光計測するため非効率的であった．本法は非標的細胞を抗原固定化微粒子でラベル化し，標的細胞のみをアレイ化できるため，千ウェル程度で標的細胞を濃縮してアレイ化可能である．また，ウェルアレイを用いた高効率細胞融合，簡便で高スループットな捕捉細胞の選択的回収を可能にしたことから，迅速，簡便，高効率なハイブリドーマ形成法を構築できる．
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