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Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked three-dimensional hydrogels


Although cell–matrix adhesive interactions are known to regulate stem cell differentiation, the underlying mechanisms, in particular for direct three-dimensional encapsulation within hydrogels, are poorly understood. Here, we demonstrate that in covalently crosslinked hyaluronic acid (HA) hydrogels, the differentiation of human mesenchymal stem cells (hMSCs) is directed by the generation of degradation-mediated cellular traction, independently of cell morphology or matrix mechanics. hMSCs within HA hydrogels of equivalent elastic moduli that permit (restrict) cell-mediated degradation exhibited high (low) degrees of cell spreading and high (low) tractions, and favoured osteogenesis (adipogenesis). Moreover, switching the permissive hydrogel to a restrictive state through delayed secondary crosslinking reduced further hydrogel degradation, suppressed traction, and caused a switch from osteogenesis to adipogenesis in the absence of changes to the extended cellular morphology. Furthermore, inhibiting tension-mediated signalling in the permissive environment mirrored the effects of delayed secondary crosslinking, whereas upregulating tension induced osteogenesis even in the restrictive environment.

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Figure 1: hMSC matrix interactions and fate choice within photopolymerized MeHA hydrogels.
Figure 2: Sequential crosslinking characterization and proteolytic degradation kinetics of MeMaHA hydrogels.
Figure 3: MeMaHA hydrogel structure-dependent hMSC matrix interactions and fate choice.
Figure 4: Delayed secondary crosslinking redirects hMSC matrix interactions and fate choice without altering cell shape.
Figure 5: hMSC matrix interactions and lineage commitment following pharmacologically induced changes in cytoskeletal tension.


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This work was supported by funding from a Fellowship in Science and Engineering from the David and Lucile Packard Foundation (J.A.B.), a CAREER award (J.A.B.) and Graduate Research Fellowship (S.K.) from the National Science Foundation, and grant GM74048 from the National Institutes of Health (C.S.C.). The authors would like to thank R. Marklein and C. Choi for helpful discussions and experimental assistance.

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S.K. and J.A.B. conceived the ideas and designed the experiments. S.K., M.G., W.R.L. and D.M.C. conducted the experiments and analysed the data. S.K., W.R.L., C.S.C. and J.A.B. interpreted the data and wrote the manuscript.

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Correspondence to Jason A. Burdick.

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The authors declare no competing financial interests.

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Khetan, S., Guvendiren, M., Legant, W. et al. Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked three-dimensional hydrogels. Nature Mater 12, 458–465 (2013).

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