Development of a miniaturized 3D organoid culture platform for ultra-high-throughput screening
| Autor: | Xingnan Li, Min Qui, Haian Fu, Xiulei Mo, Yuhong Du, Tingxuan Ma, Calvin J. Kuo, Qiankun Niu |
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| Rok vydání: | 2020 |
| Předmět: |
3D culture
2019-20 coronavirus outbreak Colon Computer science 384-well plate High-throughput screening Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Cell Culture Techniques Model system Computational biology AcademicSubjects/SCI01180 Imaging Three-Dimensional high-throughput screening (HTS) Genetics Organoid Humans Cell Shape Molecular Biology Cryopreservation Miniaturization Drug discovery Genetically engineered Articles Cell Biology General Medicine 1536-well plate ultra-HTS (uHTS) High-Throughput Screening Assays Organoids human colon organoids KRASG12D Human colon |
| Zdroj: | Journal of Molecular Cell Biology |
| ISSN: | 1759-4685 |
| DOI: | 10.1093/jmcb/mjaa036 |
| Popis: | The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery. However, organoid culturing with extracellular matrix to support 3D architecture has been challenging for high-throughput screening (HTS)-based drug discovery due to technical difficulties. Using genetically engineered human colon organoids as a model system, here we report our effort to miniaturize such 3D organoid culture with extracellular matrix support in high-density plates to enable HTS. We first established organoid culturing in a 384-well plate format and validated its application in a cell viability HTS assay by screening a 2036-compound library. We further miniaturized the 3D organoid culturing in a 1536-well ultra-HTS format and demonstrated its robust performance for large-scale primary compound screening. Our miniaturized organoid culturing method may be adapted to other types of organoids. By leveraging the power of 3D organoid culture in a high-density plate format, we provide a physiologically relevant screening platform to model tumors to accelerate organoid-based research and drug discovery. |
| Databáze: | OpenAIRE |
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