Single cell-laden protease-sensitive microniches for long-term culture in 3D.
Single cell-laden protease-sensitive microniches for long-term culture in 3D.
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DOI：
10.1039/c6lc01444e
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发表时间：
2017-02-14
期刊：
影响因子：
6.1
通讯作者：
Mooney DJ
中科院分区：
文献类型：
--
作者：
Lienemann PS;Rossow T;Mao AS;Vallmajo-Martin Q;Ehrbar M;Mooney DJ
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. We present a powerful strategy for evading Poisson encapsulation statistics and for cell centering in single cell-laden synthetic microniches to facilitate long-term culture in protease-sensitive 3D microenvironments.
