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Issue 4, 2017
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Single cell-laden protease-sensitive microniches for long-term culture in 3D

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Abstract

Single cell-laden three-dimensional (3D) microgels that can serve to mimic stem cell niches in vitro, and are therefore termed microniches, can be efficiently fabricated by droplet-based microfluidics. In this technique an aqueous polymer solution along with a highly diluted cell solution is injected into a microfluidic device to create monodisperse pre-microgel droplets that are then solidified by a polymer crosslinking reaction to obtain monodisperse single cell-laden microniches. However, problems limiting this approach studying the fate of single cells include Poisson encapsulation statistics that result in mostly empty microniches, and cells egressing from the microniches during subsequent cell culture. Here, we present a strategy to bypass Poisson encapsulation statistics in synthetic microniches by selective crosslinking of only cell-laden pre-microgel droplets. Furthermore, we show that we can position cells in the center of the microniches, and that even in protease-sensitive microniches this greatly reduces cell egress. Collectively, we present the development of a versatile protocol that allows for unprecedented efficiency in creation of synthetic protease-sensitive microniches for probing single stem cell fate in 3D.

Graphical abstract: Single cell-laden protease-sensitive microniches for long-term culture in 3D

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Publication details

The article was received on 24 Nov 2016, accepted on 25 Jan 2017 and first published on 26 Jan 2017


Article type: Paper
DOI: 10.1039/C6LC01444E
Citation: Lab Chip, 2017,17, 727-737
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    Single cell-laden protease-sensitive microniches for long-term culture in 3D

    P. S. Lienemann, T. Rossow, A. S. Mao, Q. Vallmajo-Martin, M. Ehrbar and D. J. Mooney, Lab Chip, 2017, 17, 727
    DOI: 10.1039/C6LC01444E

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