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Topics {✒️}

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         headline:Assay establishment and validation of a high-throughput organoid-based drug screening platform
         description:Organoids are three-dimensional structures that closely recapitulate tissue architecture and cellular composition, thereby holding great promise for organoid-based drug screening. Although growing in three-dimensional provides the possibility for organoids to recapitulate main features of corresponding tissues, it makes it incommodious for imaging organoids in two-dimensional and identifying surviving organoids from surrounding dead cells after organoids being treated by irradiation or chemotherapy. Therefore, significant work remains to establish high-quality controls to standardize organoid analyses and make organoid models more reproducible. In this study, the Z-stack imaging technique was used for the imaging of three-dimensional organoids to gather all the organoids’ maximum cross sections in one imaging. The combination of live cell staining fluorescent dye Calcein-AM and ImageJ assessment was used to analyze the survival of organoids treated by irradiation or chemotherapy. We have established a novel quantitative high-throughput imaging assay that harnesses the scalability of organoid cultures. Using this assay, we can capture organoid growth over time, measure multiple whole-well organoid readouts, and show the different responses to drug treatments. In summary, combining the Z-stack imaging technique and fluorescent labeling methods, we established an assay for the imaging and analysis of three-dimensional organoids. Our data demonstrated the feasibility of using organoid-based platforms for high-throughput drug screening assays.
         datePublished:2022-05-26T00:00:00Z
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      headline:Assay establishment and validation of a high-throughput organoid-based drug screening platform
      description:Organoids are three-dimensional structures that closely recapitulate tissue architecture and cellular composition, thereby holding great promise for organoid-based drug screening. Although growing in three-dimensional provides the possibility for organoids to recapitulate main features of corresponding tissues, it makes it incommodious for imaging organoids in two-dimensional and identifying surviving organoids from surrounding dead cells after organoids being treated by irradiation or chemotherapy. Therefore, significant work remains to establish high-quality controls to standardize organoid analyses and make organoid models more reproducible. In this study, the Z-stack imaging technique was used for the imaging of three-dimensional organoids to gather all the organoids’ maximum cross sections in one imaging. The combination of live cell staining fluorescent dye Calcein-AM and ImageJ assessment was used to analyze the survival of organoids treated by irradiation or chemotherapy. We have established a novel quantitative high-throughput imaging assay that harnesses the scalability of organoid cultures. Using this assay, we can capture organoid growth over time, measure multiple whole-well organoid readouts, and show the different responses to drug treatments. In summary, combining the Z-stack imaging technique and fluorescent labeling methods, we established an assay for the imaging and analysis of three-dimensional organoids. Our data demonstrated the feasibility of using organoid-based platforms for high-throughput drug screening assays.
      datePublished:2022-05-26T00:00:00Z
      dateModified:2022-05-26T00:00:00Z
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         Z-stack
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         Regenerative Medicine/Tissue Engineering
         Biomedical Engineering and Bioengineering
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         name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
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      address:
         name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
         type:PostalAddress
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      address:
         name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
         type:PostalAddress
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         name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
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      name:Xiaomeng Li
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            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
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      name:Guoxiang Fu
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            address:
               name:D1 Medical Technology Company, Shanghai, China
               type:PostalAddress
            type:Organization
      name:Long Zhang
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            address:
               name:Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
            name:Fudan University
            address:
               name:Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
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            name:Fudan University
            address:
               name:Cancer Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
      name:Ruoyu Guan
      affiliation:
            name:Fudan University
            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
      name:Peiyuan Tang
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            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
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      name:Jialing Zhang
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            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
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      name:Xinxin Rao
      affiliation:
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            address:
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      name:Shengzhi Chen
      affiliation:
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            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
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            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
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            address:
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      name:Yun Deng
      affiliation:
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            address:
               name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
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      name:Tao Lv
      affiliation:
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            address:
               name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
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      name:Xingfeng He
      affiliation:
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            address:
               name:Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
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            name:Fudan University
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               type:PostalAddress
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            address:
               name:Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
            name:Fudan University
            address:
               name:Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
      name:Peiyuan Mu
      affiliation:
            name:Fudan University
            address:
               name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
      name:Jianjun Gao
      affiliation:
            name:Fudan University
            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
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      name:Guoqiang Hua
      url:http://orcid.org/0000-0001-9274-8136
      affiliation:
            name:Fudan University
            address:
               name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
            name:Fudan University
            address:
               name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
            name:Fudan University
            address:
               name:Cancer Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
               type:PostalAddress
            type:Organization
      email:[email protected]
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      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:D1 Medical Technology Company, Shanghai, China
      name:Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China
      name:Cancer Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China
      name:Department of Colorectal Surgery, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China
      name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Institute of Radiation Medicine, Shanghai Medical College, Fudan University, Shanghai, China
      name:Department of Radiation Oncology, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China
      name:Cancer Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, China

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