Letter | Published:

Designer matrices for intestinal stem cell and organoid culture

Nature volume 539, pages 560564 (24 November 2016) | Download Citation

Abstract

Epithelial organoids recapitulate multiple aspects of real organs, making them promising models of organ development, function and disease1,2,3. However, the full potential of organoids in research and therapy has remained unrealized, owing to the poorly defined animal-derived matrices in which they are grown4. Here we used modular synthetic hydrogel networks5,6 to define the key extracellular matrix (ECM) parameters that govern intestinal stem cell (ISC) expansion and organoid formation, and show that separate stages of the process require different mechanical environments and ECM components. In particular, fibronectin-based adhesion was sufficient for ISC survival and proliferation. High matrix stiffness significantly enhanced ISC expansion through a yes-associated protein 1 (YAP)-dependent mechanism. ISC differentiation and organoid formation, on the other hand, required a soft matrix and laminin-based adhesion. We used these insights to build a fully defined culture system for the expansion of mouse and human ISCs. We also produced mechanically dynamic matrices that were initially optimal for ISC expansion and subsequently permissive to differentiation and intestinal organoid formation, thus creating well-defined alternatives to animal-derived matrices for the culture of mouse and human stem-cell-derived organoids. Our approach overcomes multiple limitations of current organoid cultures and greatly expands their applicability in basic and clinical research. The principles presented here can be extended to identify designer matrices that are optimal for long-term culture of other types of stem cells and organoids.

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Acknowledgements

We thank M. Knobloch (University of Zurich) for helpful discussions, A. Negro (EPFL) for help in the development of PEG–alginate hybrid hydrogels, D. Ossipov (Uppsala University) for providing hyaluronic acid, the Lausanne Genomic Technologies Facility (K. Harshman) for RNA-seq and D. Pioletti (EPFL) for rheometer use. N.G. was supported by an EMBO Long-Term Postdoctoral Fellowship. This work was also supported by funding from the Ecole Polytechnique Fédérale de Lausanne (EPFL). Work performed in the laboratory of H.C. was supported by the NWO Translational Adult Stem Cell Research Grant 40-41400-98-1108 and by a NWO VENI Grant 916.15.182.

Author information

Affiliations

  1. Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, School of Life Sciences (SV) and School of Engineering (STI), Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.

    • Nikolce Gjorevski
    • , Andrea Manfrin
    • , Sonja Giger
    • , Maiia E. Bragina
    •  & Matthias P. Lutolf
  2. Hubrecht Institute and University Medical Center Utrecht, Uppsalalaan 8, 3584CT Utrecht, The Netherlands.

    • Norman Sachs
    •  & Hans Clevers
  3. Swiss Institute for Experimental Cancer Research (ISREC), EPFL, Lausanne, Switzerland.

    • Paloma Ordóñez-Morán
  4. Institute of Chemical Sciences and Engineering, School of Basic Science, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.

    • Matthias P. Lutolf

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Contributions

N.G. and M.P.L. conceived the study, designed experiments, analysed data and wrote the manuscript. N.G. was involved in performing and analysing all experiments in the manuscript except for those involving human organoids. P.O.M. helped design experiments and analyse RNA-seq data. A.M. performed qPCR gene expression experiments and analysed data and produced lentiviruses. S.G. performed flow cytometry analysis of integrin expression of ISCs culture in Matrigel and PEG RGD. M.E.B. designed and characterized PEG–alg hydrogel system and helped N.G. perform experiments with ISCs in these matrices. N.S. and H.C. designed experiments and analysed data with human cells. N.S. performed experiments with human cells.

Competing interests

Ecole Polytechnique Fédérale de Lausanne (with M.P.L. and N.G.) has filed patent applications pertaining to synthetic gels for epithelial stem cell and organoid cultures. H.C. is an inventor of several patents on organoid technology.

Corresponding author

Correspondence to Matthias P. Lutolf.

Reviewer Information

Nature thanks L. Li, J. Mills and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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DOI

https://doi.org/10.1038/nature20168

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