Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Programmed assembly of synthetic protocells into thermoresponsive prototissues

Abstract

Although several new types of synthetic cell-like entities are now available, their structural integration into spatially interlinked prototissues that communicate and display coordinated functions remains a considerable challenge. Here we describe the programmed assembly of synthetic prototissue constructs based on the bio-orthogonal adhesion of a spatially confined binary community of protein–polymer protocells, termed proteinosomes. The thermoresponsive properties of the interlinked proteinosomes are used collectively to generate prototissue spheroids capable of reversible contractions that can be enzymatically modulated and exploited for mechanochemical transduction. Overall, our methodology opens up a route to the fabrication of artificial tissue-like materials capable of collective behaviours, and addresses important emerging challenges in bottom-up synthetic biology and bioinspired engineering.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Programmed assembly of proteinosomes into synthetic prototissue spheroids.
Fig. 2: Collective contractile behaviour in prototissue spheroids.
Fig. 3: Enzyme-mediated amplitude modulation within thermoresponsive prototissue spheroids.
Fig. 4: Mechanochemical transduction within prototissue spheroids.

Similar content being viewed by others

Data availability

The authors declare that all relevant data supporting the findings of this study are available within the paper and its Supplementary Information files. Additional data are available from the corresponding author upon reasonable request.

References

  1. Mantri, S. & Sapra, K. T. Evolving protocells to prototissues: rational design of a missing link. Biochem. Soc. Trans. 41, 1159–1165 (2013).

    Article  CAS  Google Scholar 

  2. Lentini, R. et al. Two-way chemical communication between artificial and natural cells. ACS Cent. Sci. 3, 117–123 (2017).

    Article  CAS  Google Scholar 

  3. Qiao, H. et al. Encapsulation of nucleic acids into giant unilamellar vesicles by freeze–thaw: a way protocells may form. Orig. Life Evol. Biosph. 47, 499–510 (2016).

    Article  Google Scholar 

  4. Altamura, E. et al. Highly oriented photosynthetic reaction centers generate a proton gradient in synthetic protocells. Proc. Natl Acad. Sci. USA 114, 3837–3842 (2017).

    Article  CAS  Google Scholar 

  5. Adamala, K. P., Engelhart, A. E. & Szostak, J. W. Collaboration between primitive cell membranes and soluble catalysts. Nat. Commun. 7, 11041 (2016).

    Article  CAS  Google Scholar 

  6. Jang, W. S. et al. Enzymatically triggered rupture of polymersomes. Soft Matter 12, 1014–1020 (2016).

    Article  CAS  Google Scholar 

  7. Peyret, A. et al. Polymersome popping by light-induced osmotic shock under temporal, spatial, and spectral control. Angew. Chem. Int. Ed. 56, 1566–1570 (2017).

    Article  CAS  Google Scholar 

  8. Schmitt, C., Lippert, A. H., Bonakdar, N., Sandoghdar, V. & Voll, L. M. Compartmentalization and transport in synthetic vesicles. Front. Bioeng. Biotechnol. 4, 19 (2016).

    Article  Google Scholar 

  9. Che, H. & van Hest, J. C. M. Stimuli-responsive polymersomes and nanoreactors. J. Mater. Chem. B 4, 4632–4647 (2016).

    Article  CAS  Google Scholar 

  10. Bellomo, E. G., Wyrsta, M. D., Pakstis, L., Pochan, D. J. & Deming, T. J. Stimuli-responsive polypeptide vesicles by conformation-specific assembly. Nat. Mater. 3, 244–248 (2004).

    Article  CAS  Google Scholar 

  11. Holowka, E. P., Sun, V. Z., Kamei, D. T. & Deming, T. J. Polyarginine segments in block copolypeptides drive both vesicular assembly and intracellular delivery. Nat. Mater. 6, 52–57 (2007).

    Article  CAS  Google Scholar 

  12. Xiao, Q. et al. Bioactive cell-like hybrids coassembled from (glyco)dendrimersomes with bacterial membranes. Proc. Natl Acad. Sci. USA 113, E1134–E1141 (2016).

    Article  CAS  Google Scholar 

  13. Percec, V. et al. Self-assembly of Janus dendrimers into uniform dendrimersomes and other complex architectures. Science 328, 1009–1014 (2010).

    Article  CAS  Google Scholar 

  14. Akkarachaneeyakorn, K., Li, M., Davis, S. A. & Mann, S. Secretion and reversible assembly of extracellular-like matrix by enzyme-active colloidosome-based protocells. Langmuir 32, 2912–2919 (2016).

    Article  CAS  Google Scholar 

  15. Sun, S. et al. Chemical signaling and functional activation in colloidosome-based protocells. Small 12, 1920–1927 (2016).

    Article  CAS  Google Scholar 

  16. Li, M., Huang, X. & Mann, S. Spontaneous growth and division in self-reproducing inorganic colloidosomes. Small 10, 3291–3298 (2014).

    Article  CAS  Google Scholar 

  17. Li, M., Harbron, R. L., Weaver, J. V., Binks, B. P. & Mann, S. Electrostatically gated membrane permeability in inorganic protocells. Nat. Chem. 5, 529–536 (2013).

    Article  CAS  Google Scholar 

  18. Yin, Y. et al. Non-equilibrium behaviour in coacervate-based protocells under electric-field-induced excitation. Nat. Commun. 7, 10658 (2016).

    Article  CAS  Google Scholar 

  19. Williams, D. S., Patil, A. J. & Mann, S. Spontaneous structuration in coacervate-based protocells by polyoxometalate-mediated membrane assembly. Small 10, 1830–1840 (2014).

    Article  CAS  Google Scholar 

  20. Tang, D. T.-Y. et al. Fatty acid membrane assembly on coacervate microdroplets as a step towards a hybrid protocell model. Nat. Chem. 6, 527–533 (2014).

    Article  Google Scholar 

  21. Tang, D. T.-Y., van Swaay, D., deMello, A., Ross Anderson, J. L. & Mann, S. In vitro gene expression within membrane-free coacervate protocells. Chem. Commun. 51, 11429–11432 (2015).

    Article  Google Scholar 

  22. Qiao, Y., Li, M., Booth, R. & Mann, S. Predatory behaviour in synthetic protocell communities. Nat. Chem. 9, 110–119 (2017).

    Article  CAS  Google Scholar 

  23. Huang, X. et al. Interfacial assembly of protein–polymer nano-conjugates into stimulus-responsive biomimetic protocells. Nat. Commun. 4, 2239 (2013).

    Article  Google Scholar 

  24. Huang, X., Li, M. & Mann, S. Membrane-mediated cascade reactions by enzyme-polymer proteinosomes. Chem. Commun. 50, 6278–6280 (2014).

    Article  CAS  Google Scholar 

  25. Huang, X., Patil, A. J., Li, M. & Mann, S. Design and construction of higher-order structure and function in proteinosome-based protocells. J. Am. Chem. Soc. 136, 9225–9234 (2014).

    Article  CAS  Google Scholar 

  26. Liu, X. et al. Hierarchical proteinosomes for programmed release of multiple components. Angew. Chem. Int. Ed. 55, 7095–7100 (2016).

    Article  CAS  Google Scholar 

  27. Wen, P. et al. Coordinated membrane fusion of proteinosomes by contact-induced hydrogel self-healing. Small 13, 1700467 (2017).

    Article  Google Scholar 

  28. Jin, H. et al. Reversible and large-scale cytomimetic vesicle aggregation: light-responsive host–guest interactions. Angew. Chem. Int. Ed. 50, 10352–10356 (2011).

    Article  CAS  Google Scholar 

  29. Jin, H. et al. Cytomimetic large-scale vesicle aggregation and fusion based on host–guest interaction. Langmuir 28, 2066–2072 (2012).

    Article  CAS  Google Scholar 

  30. Jin, H., Huang, W., Zheng, Y., Zhou, Y. & Yan, D. Construction of macroscopic cytomimetic vesicle aggregates based on click chemistry: controllable vesicle fusion and phase separation. Chemistry 18, 8641–8646 (2012).

    Article  CAS  Google Scholar 

  31. Jin, H. et al. Three-component vesicle aggregation driven by adhesion interactions between Au nanoparticles and polydopamine-coated nanotubes. Chem. Commun. 50, 6157–6160 (2014).

    Article  CAS  Google Scholar 

  32. Bai, Y. et al. A double droplet trap system for studying mass transport across a droplet–droplet interface. Lab. Chip 10, 1281–1285 (2010).

    Article  CAS  Google Scholar 

  33. Zagnoni, M. & Cooper, J. M. A microdroplet-based shift register. Lab. Chip 10, 3069–3073 (2010).

    Article  CAS  Google Scholar 

  34. Stanley, C. E. et al. A microfluidic approach for high-throughput droplet interface bilayer (DIB) formation. Chem. Commun. 46, 1620–1622 (2010).

    Article  CAS  Google Scholar 

  35. Elani, Y., Law, R. V. & Ces, O. Vesicle-based artificial cells as chemical microreactors with spatially segregated reaction pathways. Nat. Commun. 5, 5305 (2014).

    Article  CAS  Google Scholar 

  36. Dixit, S. S., Kim, H., Vasilyev, A., Eid, A. & Faris, G. W. Light-driven formation and rupture of droplet bilayers. Langmuir 26, 6193–6200 (2010).

    Article  CAS  Google Scholar 

  37. Holden, M. A., Needham, D. & Bayley, H. Functional bionetworks from nanoliter water droplets. J. Am. Chem. Soc. 129, 8650–8655 (2007).

    Article  CAS  Google Scholar 

  38. Leptihn, S. et al. Constructing droplet interface bilayers from the contact of aqueous droplets in oil. Nat. Protoc. 8, 1048–1057 (2013).

    Article  CAS  Google Scholar 

  39. Hwang, W. L., Chen, M., Cronin, B., Holden, M. A. & Bayley, H. Asymmetric droplet interface bilayers. J. Am. Chem. Soc. 130, 5878–5879 (2008).

    Article  CAS  Google Scholar 

  40. Milianta, P. J., Muzzio, M., Denver, J., Cawley, G. & Lee, S. Water permeability across symmetric and asymmetric droplet interface bilayers: interaction of cholesterol sulfate with DPhPC. Langmuir 31, 12187–12196 (2015).

    Article  CAS  Google Scholar 

  41. Villar, G., Heron, A. J. & Bayley, H. Formation of droplet networks that function in aqueous environments. Nat. Nanotech. 6, 803–808 (2011).

    Article  CAS  Google Scholar 

  42. Booth, M. J., Schild, V. R., Graham, A. D., Olof, S. N. & Bayley, H. Light-activated communication in synthetic tissues. Sci. Adv. 2, e1600056 (2016).

    Article  Google Scholar 

  43. Villar, G., Graham, A. D. & Bayley, H. A tissue-like printed material. Science 340, 48–52 (2013).

    Article  CAS  Google Scholar 

  44. Gobbo, P., Novoa, S., Biesinger, M. C. & Workentin, M. S. Interfacial strain-promoted alkyne-azide cycloaddition (I-SPAAC) for the synthesis of nanomaterial hybrids. Chem. Commun. 49, 3982–3984 (2013).

    Article  CAS  Google Scholar 

  45. Gobbo, P. et al. Versatile strained alkyne modified water-soluble AuNPs for interfacial strain promoted azide-alkyne cycloaddition (I-SPAAC). J. Mater. Chem. B 2, 1764–1769 (2014).

    Article  CAS  Google Scholar 

  46. Luo, W. et al. ‘Shine & click’ photo-induced interfacial unmasking of strained alkynes on small water-soluble gold nanoparticles. Chemistry 23, 1052–1059 (2017).

    Article  CAS  Google Scholar 

  47. Gibson, L. J. & Ashby, M. F. Cellular Solids—Structure and Properties (Cambridge Univ. Press, Cambridge, 2014).

  48. Liu, J., Sun, N., Bruce, M. A., Wu, J. C. & Butte, M. J. Atomic force mechanobiology of pluripotent stem cell-derived cardiomyocytes. PLoS ONE 7, e37559 (2012).

    Article  CAS  Google Scholar 

  49. du Roure, O. et al. Force mapping in epithelial cell migration. Proc. Natl Acad. Sci. USA 102, 2390–2395 (2005).

    Article  Google Scholar 

  50. Sabass, B., Koch, M. D., Liu, G., Stone, H. A. & Shaevitz, J. W. Force generation by groups of migrating bacteria. Proc. Natl Acad. Sci. USA 114, 7266–7271 (2017).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors thank NSERC Canada (PDF-487171–2016) and EU Horizon 2020 (Marie Skłodowska-Curie grant no. 701876) for funding, and the NMR/Mass Spectrometry Facility, the Wolfson Bioimaging Facility (A. Leard; BBSRC grant no. BB/L014181/1) and Chemical Imaging Facility (EPSRC grant ‘Atoms to Applications’, EP/K035746/1) for help with physical characterization. The authors also thank I. Manners, J. Finnegan and S. Briggs for assistance with GPC/DSC measurements, D. Woolfson for use of dichroism and plate reader spectrometers (BBSRC/EPSRC Bristol Synthetic Biology Research Centre, grant no. BB/L01386X/1), D. Gubala for help with AFM measurements, N. Martin and R. Booth for fruitful discussions, T. Ferrugia for assistance with the development of a customized microscope heating stage, and T. Liverpool for mathematical discussions.

Author information

Authors and Affiliations

Authors

Contributions

P.G., A.J.P., M.L. and S.M. conceived the experiments. P.G. performed the experiments. P.G. and R.H. performed the force measurements. W.H.B. and P.G. developed the theoretical model for the prototissue expansion force. All authors undertook data analysis, discussed the results, and contributed to drafts of the manuscript. P.G. and S.M. wrote the final manuscript.

Corresponding author

Correspondence to Stephen Mann.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Video Legends 1–5, Supplementary Notes 1–3, Supplementary Figures 1–34, Supplementary Table 1, Supplementary References 1–2

Reporting Summary

Supplementary Video 1

Caged prototissue spheroid

Supplementary Video 2

Uncaged prototissue spheroid

Supplementary Video 3

Prototissue spheroid undergoing reversible contractions

Supplementary Video 4

Prototissue undergoing reversible contractions with buckling protocells

Supplementary Video 5

Communication between compartments in an uncaged prototissue spheroid

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gobbo, P., Patil, A.J., Li, M. et al. Programmed assembly of synthetic protocells into thermoresponsive prototissues. Nature Mater 17, 1145–1153 (2018). https://doi.org/10.1038/s41563-018-0183-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41563-018-0183-5

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing