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ABSTRACT Existing techniques to encapsulate cells into microscale hydrogels generally yield high polymer-to-cell ratios and lack control over the hydrogel’s mechanical properties1. Here, we

report a microfluidic-based method for encapsulating single cells in an approximately six-micrometre layer of alginate that increases the proportion of cell-containing microgels by a factor

of ten, with encapsulation efficiencies over 90%. We show that _in vitro_ cell viability was maintained over a three-day period, that the microgels are mechanically tractable, and that, for

microscale cell assemblages of encapsulated marrow stromal cells cultured in microwells, osteogenic differentiation of encapsulated cells depends on gel stiffness and cell density. We also

show that intravenous injection of singly encapsulated marrow stromal cells into mice delays clearance kinetics and sustains donor-derived soluble factors _in vivo_. The encapsulation of

single cells in tunable hydrogels should find use in a variety of tissue engineering and regenerative medicine applications. Access through your institution Buy or subscribe This is a

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PROGRAMMED SPATIOTEMPORAL MECHANICAL CUES FOR STEM CELL-ASSISTED BONE REGENERATION Article Open access 16 April 2025 MICROINTERFACES IN BIOPOLYMER-BASED BICONTINUOUS HYDROGELS GUIDE RAPID

3D CELL MIGRATION Article Open access 29 March 2024 ENHANCED MECHANOSENSING OF CELLS IN SYNTHETIC 3D MATRIX WITH CONTROLLED BIOPHYSICAL DYNAMICS Article Open access 10 June 2021 REFERENCES *

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  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by the National Institutes of Health (NIH) Grants RO1EB014703 (D.J.M. and D.A.W.) and K99HL125884 (J.-W.S.),

and the National Science Foundation (NSF) Graduate Research Fellowship Program (A.S.M.). S.U. was supported by the Deutsche Forschungsgemeinschaft (DFG). AUTHOR INFORMATION Author notes *

Angelo S. Mao and Jae-Won Shin: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Wyss Institute for Biologically Inspired Engineering at Harvard University,

Cambridge, Massachusetts 02138, USA Angelo S. Mao, Jae-Won Shin, Oktay Uzun, Weiwei Li, David A. Weitz & David J. Mooney * School of Engineering and Applied Sciences, Harvard University,

Cambridge, Massachusetts 02138, USA Angelo S. Mao, Jae-Won Shin, Stefanie Utech, Huanan Wang, Oktay Uzun, Weiwei Li, Madeline Cooper, Yuebi Hu, Liyuan Zhang, David A. Weitz & David J.

Mooney * Department of Pharmacology and Department of Bioengineering, University of Illinois College of Medicine, Chicago, Illinois 60612, USA Jae-Won Shin * Department of Physics, Harvard

University, Cambridge, Massachusetts 02138, USA Huanan Wang, Liyuan Zhang & David A. Weitz * Biomaterials and Tissue Engineering Laboratory, School of Life Science and Biotechnology,

Dalian University of Technology, Dalian 116024, China Huanan Wang Authors * Angelo S. Mao View author publications You can also search for this author inPubMed Google Scholar * Jae-Won Shin

View author publications You can also search for this author inPubMed Google Scholar * Stefanie Utech View author publications You can also search for this author inPubMed Google Scholar *

Huanan Wang View author publications You can also search for this author inPubMed Google Scholar * Oktay Uzun View author publications You can also search for this author inPubMed Google

Scholar * Weiwei Li View author publications You can also search for this author inPubMed Google Scholar * Madeline Cooper View author publications You can also search for this author

inPubMed Google Scholar * Yuebi Hu View author publications You can also search for this author inPubMed Google Scholar * Liyuan Zhang View author publications You can also search for this

author inPubMed Google Scholar * David A. Weitz View author publications You can also search for this author inPubMed Google Scholar * David J. Mooney View author publications You can also

search for this author inPubMed Google Scholar CONTRIBUTIONS A.S.M., J.-W.S. and D.J.M. conceived and designed the experiments. S.U., H.W. and D.A.W. contributed to microfluidic design and

fabrication. A.S.M. and J.-W.S. performed the experiments. A.S.M., J.-W.S. and D.J.M. analysed the data. A.S.M., J.-W.S. and D.J.M. wrote the manuscript. All authors discussed the results

and commented on the manuscript. A.S.M. and J.-W.S. contributed equally to this work. The principal investigator is D.J.M. CORRESPONDING AUTHOR Correspondence to David J. Mooney. ETHICS

DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary Information (PDF 2659 kb) RIGHTS AND

PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Mao, A., Shin, JW., Utech, S. _et al._ Deterministic encapsulation of single cells in thin tunable microgels for

niche modelling and therapeutic delivery. _Nature Mater_ 16, 236–243 (2017). https://doi.org/10.1038/nmat4781 Download citation * Received: 04 October 2015 * Accepted: 22 September 2016 *

Published: 31 October 2016 * Issue Date: February 2017 * DOI: https://doi.org/10.1038/nmat4781 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content:

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