Microfluidic platform for live cell imaging of 3D cultures with clone retrieval
| Autor: | Carla Mulas, Chibeza C. Agley, Florian Hollfelder, Andrew C Hodgson, Timo N Kohler, Kevin J. Chalut, Austin Smith |
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| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Computer science 030303 biophysics Microfluidics Cell Embryonic stem cell Cell biology 03 medical and health sciences medicine.anatomical_structure Live cell imaging medicine Cellular dynamics Cell encapsulation Clone (B-cell biology) Combined method 030304 developmental biology |
| Zdroj: | bioRxiv |
| DOI: | 10.1101/2020.02.17.952689 |
| Popis: | Combining live imaging with the ability to retrieve individual cells of interest remains a technical challenge. These combined methods are of particular interest when studying highly dynamic or transient, asynchronous or heterogeneous cell biological and developmental processes. Here we present a method to encapsulate live cells in a 3D hydrogel matrix, via droplet compartmentalisation. Using a small-scale screen, we optimised matrix conditions for the culture and multilineage differentiation of mouse embryonic stem (ES) cells. Moreover, we designed a custom microfluidic platform that is compatible with live imaging. With this platform we are able to retain or extract individual bead/droplets by media flow only, obviating the need for enzymatic cell removal from the platform. We show that we can differentiate mES cells, monitor reporter expression by live imaging, and retrieve individual droplets for functional assays, correlating reporter expression with functional response. Overall, we present a highly flexible 3D cell encapsulation and microfluidic platform that enables both monitoring of cellular dynamics and retrieval for molecular and functional assays. |
| Databáze: | OpenAIRE |
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