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In situ collagen assembly for integrating microfabricated three-dimensional cell-seeded matrices

Nature Materials volume 7, pages 636–640 (2008)Cite this article

Abstract

Microscale fabrication of three-dimensional (3D) extracellular matrices (ECMs) can be used to mimic the often inhomogeneous and anisotropic properties of native tissues1,2,3 and to construct in vitro cellular microenvironments4,5,6. Cellular contraction of fibrous natural ECMs (such as fibrin and collagen I) can detach matrices from their surroundings and destroy intended geometry7,8,9. Here, we demonstrate in situ collagen fibre assembly (the nucleation and growth of new collagen fibres from preformed collagen fibres at an interface) to anchor together multiple phases of cell-seeded 3D hydrogel-based matrices against cellular contractile forces. We apply this technique to stably interface multiple microfabricated 3D natural matrices (containing collagen I, Matrigel, fibrin or alginate); each phase can be seeded with cells and designed to permit cell spreading. With collagen-fibre-mediated interfacing, microfabricated 3D matrices maintain stable interfaces (the individual phases do not separate from each other) over long-term culture (at least 3 weeks) and support spatially restricted development of multicellular structures within designed patterns. The technique enables construction of well-defined and stable patterns of a variety of 3D ECMs formed by diverse mechanisms (including temperature-, ion- and enzyme-mediated crosslinking), and presents a simple approach to interface multiple 3D matrices for biological studies and tissue engineering.

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Figure 1: Use and characterization of in situ assembly of collagen fibres to integrate multiple 3D matrices.
Figure 2: Imaging of micropatterned HUVEC-seeded collagen within bulk hydrogel scaffolds.
Figure 3: Stabilization of patterned ECM by interfacing collagen fibres against cellular contractile forces.
Figure 4: Cellular morphology and behaviour in bulk and patterned phases.

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Acknowledgements

We acknowledge a Scientist Development Grant from the American Heart Association and an NSF CAREER award.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 1210 Amsterdam Avenue, New York 10027, USA

    Brian M. Gillette, Jacob A. Jensen, Beixian Tang, Genevieve J. Yang, Ardalan Bazargan-Lari, Ming Zhong & Samuel K. Sia

Authors
  1. Brian M. Gillette
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  2. Jacob A. Jensen
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  3. Beixian Tang
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  4. Genevieve J. Yang
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  5. Ardalan Bazargan-Lari
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  6. Ming Zhong
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  7. Samuel K. Sia
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Contributions

B.M.G. and S.K.S. conceived the project, designed the experiments, analysed the data, developed the interpretations and wrote the manuscript. B.M.G. carried out the bulk of the experiments. J.A.J., B.T., G.J.Y., A.B. and M.Z. provided suggestions for developing the method and carried out the remainder of the experiments under the guidance of B.M.G. and S.K.S. All authors have agreed to the content of the manuscript, including the data as presented.

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Correspondence to Samuel K. Sia.

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Gillette, B., Jensen, J., Tang, B. et al. In situ collagen assembly for integrating microfabricated three-dimensional cell-seeded matrices. Nature Mater 7, 636–640 (2008). https://doi.org/10.1038/nmat2203

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