Article

Synthetic hydrogels for human intestinal organoid generation and colonic wound repair

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Abstract

In vitro differentiation of human intestinal organoids (HIOs) from pluripotent stem cells is an unparalleled system for creating complex, multicellular three-dimensional structures capable of giving rise to tissue analogous to native human tissue. Current methods for generating HIOs rely on growth in an undefined tumour-derived extracellular matrix (ECM), which severely limits the use of organoid technologies for regenerative and translational medicine. Here, we developed a fully defined, synthetic hydrogel based on a four-armed, maleimide-terminated poly(ethylene glycol) macromer that supports robust and highly reproducible in vitro growth and expansion of HIOs, such that three-dimensional structures are never embedded in tumour-derived ECM. We also demonstrate that the hydrogel serves as an injection vehicle that can be delivered into injured intestinal mucosa resulting in HIO engraftment and improved colonic wound repair. Together, these studies show proof-of-concept that HIOs may be used therapeutically to treat intestinal injury.

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Acknowledgements

This research was supported by the National Institute of Health (A.J.G. was supported by R01 AR062368 and R01 AR062920; A.N. was supported by DK055679, DK059888 and DK089763) and a seed grant from the Regenerative Engineering and Medicine Research Center between Emory University, Georgia Tech and the University of Georgia. J.R.S. was supported by the Intestinal Stem Cell Consortium (U01DK103141), a collaborative research project funded by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and the National Institute of Allergy and Infectious Diseases (NIAID), by the NIAID Novel, Alternative Model Systems for Enteric Diseases (NAMSED) consortium (U19AI116482) and PHS Grant UL1TR000454 from the Clinical and Translational Science Award Program. R.C.-A. is supported by the National Science Foundation Graduate Research Fellowship (DGE-1650044) and the Alfred P. Sloan Foundation’s Minority Ph.D. (MPHD) Program (G-2016-20166039). M.Q. is supported by a fellowship from the Crohn’s and Colitis Foundation of America (CCFA 326912). A.E.F. is supported by János Bolyai Research Fellowship (BO/00023/17/8). We would like to thank Y.-H. Tsai (University of Michigan, USA) for providing tissue sections for immunohistochemical analysis.

Author information

Author notes

    • Ricardo Cruz-Acuña
    •  & Miguel Quirós

    These authors contributed equally to this work.

Affiliations

  1. Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA

    • Ricardo Cruz-Acuña
  2. Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, Georgia 30332, USA

    • Ricardo Cruz-Acuña
    •  & Andrés J. García
  3. Department of Pathology, University of Michigan, Ann Arbor, Michigan 48104, USA

    • Miguel Quirós
    • , Dorothée Siuda
    • , Vicky García-Hernández
    •  & Asma Nusrat
  4. Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged 6726, Hungary

    • Attila E. Farkas
  5. Department of Internal Medicine, Division of Gastroenterology, University of Michigan Medical School, Ann Arbor, Michigan 48104, USA

    • Priya H. Dedhia
    • , Sha Huang
    • , Alyssa J. Miller
    •  & Jason R. Spence
  6. Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, Michigan 48104, USA

    • Priya H. Dedhia
    • , Sha Huang
    •  & Jason R. Spence
  7. Center for Organogenesis, University of Michigan Medical School, Ann Arbor, Michigan 48104, USA

    • Priya H. Dedhia
    • , Sha Huang
    •  & Jason R. Spence
  8. George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA

    • Andrés J. García

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Contributions

R.C.-A. and M.Q. conducted all experiments, collected data and performed data analyses. A.E.F. assisted with in vitro and in vivo intestinal crypt experiments. P.H.D. performed the in vivo HIO implantation under the kidney capsule experiment. S.H. performed the culture and differentiation of PSCs into intestinal spheroids. D.S. assisted with in vivo HIO delivery to mouse colonic wounds. V.G.-H. and A.J.M. performed the in situ hybridization experiments. A.N., A.J.G. and J.R.S. conceptualized and designed the project and experiments. R.C.-A., A.J.G., A.N., J.R.S. and M.Q. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Jason R. Spence or Asma Nusrat or Andrés J. García.

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