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Stress-stiffening-mediated stem-cell commitment switch in soft responsive hydrogels

Nature Materials volume 15, pages 318–325 (2016)Cite this article

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Abstract

Bulk matrix stiffness has emerged as a key mechanical cue in stem cell differentiation. Here, we show that the commitment and differentiation of human mesenchymal stem cells encapsulated in physiologically soft (∼0.2–0.4 kPa), fully synthetic polyisocyanopeptide-based three-dimensional (3D) matrices that mimic the stiffness of adult stem cell niches and show biopolymer-like stress stiffening, can be readily switched from adipogenesis to osteogenesis by changing only the onset of stress stiffening. This mechanical behaviour can be tuned by simply altering the material’s polymer length whilst maintaining stiffness and ligand density. Our findings introduce stress stiffening as an important parameter that governs stem cell fate in a 3D microenvironment, and reveal a correlation between the onset of stiffening and the expression of the microtubule-associated protein DCAMKL1, thus implicating DCAMKL1 in a stress-stiffening-mediated, mechanotransduction pathway that involves microtubule dynamics in stem cell osteogenesis.

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Figure 1: Differential modulus, K’, as a function of stress, σ, for intracellular and extracellular filamentous biopolymer gels that show stress stiffening.
Figure 2: Synthesis of polymers of different chain lengths and characterization of polymer hydrogels.
Figure 3: Effect of stress stiffening on hMSC commitment.
Figure 4: Stress-stiffening-mediated stem cell differentiation involves the microtubule-associated protein DCAMKL1.

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Acknowledgements

This work was supported by NWO grant 728.011.102 (R.K.D.), Gravitation grant Functional Molecular System (A.E.R.), NanoNext grant 3D.12 (R.H.), Histide AG and Histide Lab. We acknowledge P. Kouwer for discussions on stress stiffening. We also acknowledge K. Blank for useful discussions on biofunctionalization. We also thank R. Nolte for his interest in the work and useful general suggestions.

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Author notes

  1. Rajat K. Das and Veronika Gocheva: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Molecules and Materials, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands

    Rajat K. Das, Roel Hammink & Alan E. Rowan

  2. Histide, Chaltenbodenstrasse 8, 8834 Schindellegi, Switzerland

    Veronika Gocheva & Omar F. Zouani

  3. Histide Lab, Accinov, 317, avenue Jean Jaurès, 69007 Lyon, France

    Veronika Gocheva & Omar F. Zouani

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  1. Rajat K. Das
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Contributions

R.K.D., O.F.Z. and A.E.R. conceived and initiated the project. R.K.D. and R.H. synthesized the polymers and did the rheological characterization. V.G. and O.F.Z. performed the stem cell experiments. R.H. performed the AFM experiments. O.F.Z., R.K.D., V.G., R.H. and A.E.R. analysed the data. R.K.D., V.G., A.E.R. and O.F.Z. wrote the manuscript. A.E.R. and O.F.Z. supervised the project.

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Correspondence to Rajat K. Das, Omar F. Zouani or Alan E. Rowan.

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Das, R., Gocheva, V., Hammink, R. et al. Stress-stiffening-mediated stem-cell commitment switch in soft responsive hydrogels. Nature Mater 15, 318–325 (2016). https://doi.org/10.1038/nmat4483

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