CellRaft® AIR システムによる自動セルクローニングシステム
~細胞のイメージング・識別・分離とCHO細胞への応用例~
[Webinarオンデマンド配信 ] ライブ実施日時:2021/10/28
シングルセルクローニングは単一クローンに由来した細胞株を樹立する手法であり、遺伝子導入した細胞株や抗体産生細胞の樹立などに必須の手法です。代表的なシングルセルクローニング法である限界希釈法やFACSは、大量のプレートや培地などの消耗品が必要になったり、大型かつ高価な機器の購入が必要だったり、あるいは単離後の細胞へのダメージが大きいといった問題点がありました。
Cell Microsystems社で開発されたCellRaft
® AIRシステムは、極小のウェルが無数に並んだアレイを用いることで迅速かつ高い細胞生存率(80-90%)でシングルセル化が行えます。シングルセル化後にアレイ上で細胞を培養可能であり、成長率・クローナリティ・蛍光シグナル強度(3 channel)などから細胞を識別し、任意の細胞コロニーを培養プレートやPCRチューブに素早く単離することが可能です。さらに、数万に及ぶシングルセルの増殖過程は時系列の画像データとして全てアーカイブ化されるので、時を遡って細胞の状態を確認することも可能です。この一連の作業の大部分を自動化することにより、CellRaft
® AIRシステムは極めて効率的なシングルセルのワークフローをお客様に提供します。
本セミナーではCellRaft
® AIRシステムの特長と、CHO細胞を使用したモデルケースについてご紹介します。
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【 CRISPR 】
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【 CellRaft Technology 】Automated microarray for single-cell sorting and collection of lymphocytes following HIV reactivationBelén Cortés-Llanos, Vaibhav Jain, Alicia Volkheimer, Edward P Browne, David M Murdoch, Nancy L Allbritton
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Identification and isolation of antigen-specific cytotoxic T lymphocytes with an automated microraft sorting system.Attayek PJ, Hunsucker SA, Sims CE, Allbritton NL, Armistead PM.
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Array of Biodegradable Microrafts for Isolation and Implantation of Living, Adherent CellsWang Y, Phillips CN, Herrera GS, Sims CE, Yeh JJ, Allbritton NL
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Dissolution-guided wetting for microarray and microfluidic devices.Wang Y, Sims CE, Allbritton NL
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【 Genomics 】Single-cell somatic copy number variants in brain using different amplification methods and reference genomesKalef-Ezra et al., bioRxiv Posted August 08, 2023. doi: https://doi.org/10.1101/2023.08.07.552289
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Pooled image-base screening of mitochondria with microraft isolation distinguishes pathogenic mitofusin 2 mutationsAlex L. Yenkin, John C. Bramley, Colin L. Kremitzki, Jason E. Waligorski, Mariel J. Liebeskind et al.
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Selective single cell isolation for genomics using microraft arrays.Welch JD, Williams LA, DiSalvo M, Brandt AT, Marayati R, Sims CE, Allbritton NL, Prins JF, Yeh JJ, Jones CD.
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【 Organoids 】Hypoxia Primes Human ISCs for Interleukin-Dependent Rescue of Stem Cell ActivityKristina R Rivera et al., Cell Mol Gastroenterol Hepatol. 2023;16(5):823-846. doi: 10.1016/j.jcmgh.2023.07.012
The CellRaft AIR® system: A novel system enabling organoid imaging, identification, and isolationAllysa Stern, Brandon Thompson, Keith Williams, Rob McClellan, Steven Gebhart, Jessica Hartman
SLAS Discov. 2022 Apr;27(3):201-208. doi: 10.1016/j.slasd.2021.11.003. Epub 2021 Dec 4.
A high-throughput platform for stem cell niche co-cultures and downstream gene expression analysis.Gracz AD, Williamson IA, Roche KC, Johnston MJ, Wang F, Wang Y, Attayek PJ, Balowski J, Liu XF, Laurenza RJ, Gaynor LT, Sims CE, Galanko JA, Li L, Allbritton NL, Magness ST
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【 Stem Cells 】RNA binding protein SYNCRIP maintains proteostasis and self-renewal of hematopoietic stem and progenitor cellsFlorisela Herrejon Chavez, Hanzhi Luo, Paolo Cifani, Alli Pine, Karen L. Chu et al.
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Automated sensing and splitting of stem cell colonies on microraft arraysDiSalvo M, Smiddy NM, Allbritton NL
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Transferable neuronal mini-cultures to accelerate screening in primary and induced pluripotent stem cell-derived neurons.Niedringhaus M, Dumitru R, Mabb AM, Wang Y, Philpot BD, Allbritton NL, Taylor AM
Sci Rep. 2015 Feb 10;5:8353. doi: 10.1038/srep08353
A high-throughput platform for stem cell niche co-cultures and downstream gene expression analysis.Gracz AD, Williamson IA, Roche KC, Johnston MJ, Wang F, Wang Y, Attayek PJ, Balowski J, Liu XF, Laurenza RJ, Gaynor LT, Sims CE, Galanko JA, Li L, Allbritton NL, Magness ST
Nat Cell Biol. 2015 Mar;17(3):340-9. doi: 10.1038/ncb3104. Epub 2015 Feb 9
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