Massively parallel encapsulation of single cells with structured microparticles and secretion-based flow sorting
| Autor: | Joseph de Rutte, Robert Dimatteo, Maani M Archang, Mark van Zee, Doyeon Koo, Sohyung Lee, Allison C. Sharrow, Patrick J. Krohl, Michael P. Mellody, Sheldon Zhu, James Eichenbaum, Monika Kizerwetter, Shreya Udani, Kyung Ha, Andrea L. Bertozzi, Jamie B. Spangler, Robert Damoiseaux, Dino Di Carlo |
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| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Chemistry Chinese hamster ovary cell Cell 02 engineering and technology 021001 nanoscience & nanotechnology Encapsulation (networking) Cell biology Flow sorting 03 medical and health sciences medicine.anatomical_structure medicine Secretion 0210 nano-technology Massively parallel 030304 developmental biology |
| Popis: | Techniques to analyze and sort single cells based on functional outputs, such as secreted products, have the potential to transform our understanding of cellular biology, as well as accelerate the development of next generation cell and antibody therapies. However, secreted molecules rapidly diffuse away from cells, and analysis of these products requires specialized equipment and expertise to compartmentalize individual cells and capture their secretions. Herein we demonstrate the use of suspendable microcontainers to sort single viable cells based on their secreted products at high-throughput using only commonly accessible laboratory infrastructure. Our microparticles act as solid supports which facilitate cell attachment, partition uniform aqueous compartments, and capture secreted proteins. Using this platform, we demonstrate high-throughput screening of stably- and transiently-transfected producer cells based on relative IgG production as well as screening of B lymphocytes and hybridomas based on antigen-specific antibody production using commercially available flow sorters. Leveraging the high-speed sorting capabilities of standard sorters, we sorted >1,000,000 events in less than an hour. The reported microparticles can be easily stored, and distributed as a consumable reagent amongst researchers, democratizing access to high-throughput functional cell screening. |
| Databáze: | OpenAIRE |
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