Microfluidic platform for 3D cell culture with live imaging and clone retrieval (Electronic supplementary information (ESI) available. See DOI: 10.1039/d0lc00165a)

Combining live imaging with the ability to retrieve individual cells of interest remains a technical challenge. Combining imaging with precise cell retrieval is of particular interest when studying highly dynamic or transient, asynchronous, or heterogeneous cell biological and developmental processe...

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Veröffentlicht in:Lab on a chip 2020-01, Vol.20 (14), p.2580-2591
Hauptverfasser: Mulas, Carla, Hodgson, Andrew C, Kohler, Timo N, Agley, Chibeza C, Humphreys, Peter, Kleine-Brüggeney, Hans, Hollfelder, Florian, Smith, Austin, Chalut, Kevin J
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container_end_page 2591
container_issue 14
container_start_page 2580
container_title Lab on a chip
container_volume 20
creator Mulas, Carla
Hodgson, Andrew C
Kohler, Timo N
Agley, Chibeza C
Humphreys, Peter
Kleine-Brüggeney, Hans
Hollfelder, Florian
Smith, Austin
Chalut, Kevin J
description Combining live imaging with the ability to retrieve individual cells of interest remains a technical challenge. Combining imaging with precise cell retrieval is 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 hydrogel bead compartmentalisation. Using a small-scale screen, we optimised matrix conditions for the culture and multilineage differentiation of mouse embryonic stem cells. Moreover, we designed a custom microfluidic platform that is compatible with live imaging. With this platform we can long-term culture and subsequently extract individual cells-in-beads by media flow only, obviating the need for enzymatic cell removal from the platform. Specific beads may be extracted from the platform in isolation, without disrupting the adjacent beads. We show that we can differentiate mouse embryonic stem cells, monitor reporter expression by live imaging, and retrieve individual beads 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.
doi_str_mv 10.1039/d0lc00165a
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source Royal Society Of Chemistry Journals; Alma/SFX Local Collection
subjects Beads
Biological activity
Encapsulation
Hydrogels
Imaging
Information retrieval
Microfluidics
Stem cells
title Microfluidic platform for 3D cell culture with live imaging and clone retrieval (Electronic supplementary information (ESI) available. See DOI: 10.1039/d0lc00165a)
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