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Organoids

Nature Reviews Methods Primers volume 2, Article number: 94 (2022) Cite this article

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Abstract

Organoids are simple tissue-engineered cell-based in vitro models that recapitulate many aspects of the complex structure and function of the corresponding in vivo tissue. They can be dissected and interrogated for fundamental mechanistic studies on development, regeneration and repair in human tissues, and can also be used in diagnostics, disease modelling, drug discovery and personalized medicine. Organoids are derived from either pluripotent or tissue-resident stem (embryonic or adult) or progenitor or differentiated cells from healthy or diseased tissues, such as tumours. To date, numerous organoid engineering strategies that support organoid culture and growth, proliferation, differentiation and maturation have been reported. This Primer highlights the rationale underlying the selection and development of these materials and methods to control the cellular/tissue niche; and therefore, the structure and function of the engineered organoid. We also discuss key considerations for generating robust organoids, such as those related to cell isolation and seeding, matrix and soluble factor selection, physical cues and integration. The general standards for data quality, reproducibility and deposition within the organoid community are also outlined. Lastly, we conclude by elaborating on the limitations of organoids in different applications, and the key priorities in organoid engineering for the coming years.

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Fig. 1: Components of organoid engineering.
Fig. 2: Flowchart of the procedures.
Fig. 3: Representative results of pancreatic islet organoid validation analysis.
Fig. 4: Typical characterization of cancer organoids: liver cancer subtype131.
Fig. 5: Reducing the heterogeneity with complexity reduction.
Fig. 6: Side by side comparison of the current limitations for organoid culture and approaches to overcome them.

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Acknowledgements

This work was supported in part by grants from CAS (XDA16020200 to Y.A.Z.), the National Key Research, Development Program of China (2020YFA0509002 to Y.A.Z.) and the National Institutes of Health (NIH) (R01CA240718, R01CA244729 and R01CA264248 to A.So.), and funding from the Institute of Bioengineering and Bioimaging, Biomedical Research Council, Agency for Science, Technology and Research (A*STAR) (Project Number IAF (H18/01/a0/017)), SMART CAMP, The Institute for Digital Medicine (WisDM) and Mechanobiology Institute of Singapore (R-714-106-004-135) to H.Y.

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Author notes

  1. These authors contributed equally: Zixuan Zhao, Xinyi Chen, Anna M. Dowbaj, Aleksandra Sljukic, Kaitlin Bratlie, Luda Lin.

Authors and Affiliations

  1. Mechanobiology Institute, National University of Singapore, Singapore, Singapore

    Zixuan Zhao & Hanry Yu

  2. State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, China

    Xinyi Chen, Zhaowei Chen & Yi Arial Zeng

  3. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    Anna M. Dowbaj, Aleksandra Sljukic & Meritxell Huch

  4. Department of Chemical and Biological Engineering, Iowa State University, Ames, IA, USA

    Kaitlin Bratlie & Qun Wang

  5. Department of Orthopaedic Surgery, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA

    Luda Lin & Alice Soragni

  6. Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA, USA

    Luda Lin & Alice Soragni

  7. Translational Tumor Engineering Laboratory, Department of Biomedical Engineering, National University of Singapore, Singapore, Singapore

    Eliza Li Shan Fong

  8. The N.1 Institute for Health, National University of Singapore, Singapore, Singapore

    Eliza Li Shan Fong

  9. Department of Physiology, Institute for Digital Medicine (WisDM), Yong Loo Lin School of Medicine, Singapore, Singapore

    Gowri Manohari Balachander & Hanry Yu

  10. Jonsson Comprehensive Cancer Center, University of California Los Angeles, Los Angeles, CA, USA

    Alice Soragni

  11. Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California Los Angeles, Los Angeles, CA, USA

    Alice Soragni

  12. California NanoSystems Institute, University of California Los Angeles, Los Angeles, CA, USA

    Alice Soragni

  13. School of Life Science, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Hangzhou, China

    Yi Arial Zeng

  14. Institute of Bioengineering and Bioimaging, A*STAR, Singapore, Singapore

    Hanry Yu

  15. CAMP, Singapore-MIT Alliance for Research and Technology, Singapore, Singapore

    Hanry Yu

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  1. Zixuan Zhao
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  2. Xinyi Chen
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  3. Anna M. Dowbaj
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  4. Aleksandra Sljukic
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Contributions

Introduction (H.Y., Z.Z. and E.L.S.F.); Experimentation (Z.Z., K.B., E.L.S.F. and Q.W.); Results (X.C., A.M.D., A.Sl., L.L., Z.C., A.So., M.H. and Y.A.Z.); Applications (L.L. and A.So.); Reproducibility and data deposition (A.M.D., A.Sl. and M.H.); Limitations and optimizations (H.Y., A.M.D., A.Sl. and M.H.); Outlook (H.Y., Z.Z. and G.M.B.); Overview of the Primer (H.Y.).

Corresponding authors

Correspondence to Alice Soragni, Meritxell Huch, Yi Arial Zeng, Qun Wang or Hanry Yu.

Ethics declarations

Competing interests

M.H. is inventor on several patents on organoid technology. A.So. and L.L. are inventors on a patent on organoid technology. A.So. is founder and owner of Icona BioDx. H.Y. is inventor on several patents on cell-based models and founder of Ants Innovate and Invitrocue. The remaining authors declare no competing interests.

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Nature Reviews Methods Primers thanks Sebastian Merker, Toshiro Sato, Michael Schumacher and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Glossary

2.5D culture

3D culture with a restricted third dimension.

Assembloids

Complex 3D structures combining several separately pre-generated cellular compartments and/or entities.

Bioprinting

An additive manufacturing technique that enables direct deposition of stem cells, organoids and biomaterials to fabricate 3D organoid-based tissue structures with controlled cell–matrix structures.

Engraftment rates

Degrees of organoid retention in a host tissue after transplantation.

Organogenesis

The series of organized integrated processes that transform an amorphous mass of cells into a complete organ in the developing embryo.

Passaging

The subculturing of organoids by creating either smaller organoid fragments or single cells using mechanical dissociation or enzymatic digestion.

Stem cell niche

A specific tissue microenvironment where stem cells both reside and receive stimuli that regulate cell fate.

Streptozotocin

An agent toxic to the insulin-producing β-cells of the pancreas in mammals.

Temporal heterogeneity

Here, differences in the composition and changes during the time of passaging the organoids.

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Zhao, Z., Chen, X., Dowbaj, A.M. et al. Organoids. Nat Rev Methods Primers 2, 94 (2022). https://doi.org/10.1038/s43586-022-00174-y

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