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Automated cell isolation from photodegradable hydrogel based on fluorescence image analysis
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  • Shinji Sugiura,
  • Shinya Yamahira,
  • Masato Tamura,
  • Kazumi Shin,
  • Mayu Shibuta,
  • Taku Satoh,
  • Yui Matsuzawa,
  • Gen Fujii,
  • Fumiki Yanagawa,
  • Michihiro Mutoh,
  • Masumi Yanagisawa,
  • Ryuji Kato,
  • Hirofumi Matsui
Shinji Sugiura
Sangyo Gijutsu Sogo Kenkyujo Tsukuba Higashi

Corresponding Author:shinji.sugiura@aist.go.jp

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Shinya Yamahira
Sangyo Gijutsu Sogo Kenkyujo Tsukuba Higashi
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Masato Tamura
Sangyo Gijutsu Sogo Kenkyujo Tsukuba Higashi
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Kazumi Shin
Sangyo Gijutsu Sogo Kenkyujo Tsukuba Higashi
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Mayu Shibuta
Nagoya Daigaku Fuzoku Toshokan
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Taku Satoh
Sangyo Gijutsu Sogo Kenkyujo Tsukuba Higashi
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Yui Matsuzawa
Kokuritsu Gan Kenkyu Center Gan Yobo Kenshin Kenkyu Center
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Gen Fujii
Kokuritsu Gan Kenkyu Center Gan Yobo Kenshin Kenkyu Center
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Fumiki Yanagawa
Sangyo Gijutsu Sogo Kenkyujo Tsukuba Higashi
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Michihiro Mutoh
Kokuritsu Gan Kenkyu Center Gan Yobo Kenshin Kenkyu Center
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Masumi Yanagisawa
Engineering System Co Ltd 5652-83 Sasaga Matsumoto Nagano 399-0033 Japan
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Ryuji Kato
Nagoya Daigaku Fuzoku Toshokan
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Hirofumi Matsui
Tsukuba Daigaku Igaku Iryokei
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Abstract

We report an automated cell-isolation system based on fluorescence image analysis of cell aggregates cultured in a photodegradable hydrogel. The system incorporates cell culture in a humidified atmosphere with controlled CO 2 concentration and temperature, image acquisition and analysis, micropatterned light exposure, and cell collection by pipetting. Cell aggregates were cultured on hydrogels, and target cells were selected by phase contrast and fluorescence image analysis. After degradation of the hydrogel by exposure to micropatterned ultraviolet light, cell aggregates were transferred to a collection vessel by robotic pipetting. We assessed the system for hydrogel degradation, recovery of target cells, and contamination by off-target cells. We demonstrated two practical applications of our method: (i) in cell aggregates from MCF-7-RFP strains in which 18.8% of cells produced red fluorescent protein (RFP), we successfully obtained 14 proliferative fluorescence-positive cell aggregates from 31 wells, and all of the isolated strains produced a higher proportion of RFP than the original populations; (ii) after fluorescent immunostaining of human epidermal growth factor receptor 2 (HER2) in cancer cells, we successfully isolated HER2-positive cells from a mixed population of HER2-positive and -negative cells, and gene sequence analysis confirmed that the isolated cells mainly contained the target cells.
25 Apr 2022Submitted to Biotechnology and Bioengineering
27 Apr 2022Submission Checks Completed
27 Apr 2022Assigned to Editor
30 May 2022Reviewer(s) Assigned
22 Sep 2022Review(s) Completed, Editorial Evaluation Pending
22 Sep 2022Editorial Decision: Revise Major
10 Jan 20231st Revision Received
10 Jan 2023Submission Checks Completed
10 Jan 2023Assigned to Editor
10 Jan 2023Review(s) Completed, Editorial Evaluation Pending
04 Mar 2023Reviewer(s) Assigned
12 Mar 2023Editorial Decision: Accept