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  • Review Article
  • Published:

Organoid bioprinting: from cells to functional tissues

Nature Reviews Bioengineering volume 3, pages 126–142 (2025)Cite this article

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Abstract

The biofabrication of complex human tissues to recapitulate organ-specific architecture and function requires a combination of engineering control and intrinsic self-assembly. Organoid bioprinting encompasses additive manufacturing approaches that can impart spatial control over the placement of organoids or organoid-forming cells to fabricate multicellular, 3D structures. In particular, bioprinting can be leveraged to control the spatial positioning of printed cells or tissues while maintaining the architecture and physiology of the constituent building blocks. In this Review, we discuss the emerging integration of bioprinting methods and tissue engineering. As bioprinting conventionally involves the patterning of a ‘material’ ink, we characterize cells and organoids as a living material and discuss how such a living material can be manipulated through biofabrication techniques. We focus on continuous and pick-and-place bioprinting methods in which spheroids, organoids or organoid-forming cells comprise the bioink. Additionally, we discuss organoid support baths into which inks are printed. Finally, we highlight how the combination of bioprinting approaches and organoid technology has the potential to improve engineered tissue models of development and disease.

Key points

  • Organoids are in vitro model systems designed to recapitulate in vivo human, tissue-specific phenotypes.

  • Organoid-forming cells, organoids, organoid suspensions and fused organoids can all be considered ‘living materials’, which indicates that they can be directly used in material-based biofabrication approaches.

  • Organoid bioprinting refers to additive manufacturing approaches that impart spatial control over the 3D arrangement of organoids or organoid-forming cells to increase the relevance, reproducibility and complexity of engineered tissues.

  • Organoid bioprinting holds potential for modelling human development and disease, standardizing drug discovery and development and biomanufacturing of human tissues for clinical use.

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Fig. 1: Living materials for bioprinting.
Fig. 2: Organoid bioprinting approaches.
Fig. 3: Applications for organoid bioprinting.

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References

  1. Rossi, G., Manfrin, A. & Lutolf, M. P. Progress and potential in organoid research. Nat. Rev. Genet. 19, 671–687 (2018).

    MATH  Google Scholar 

  2. Fatehullah, A., Tan, S. H. & Barker, N. Organoids as an in vitro model of human development and disease. Nat. Cell Biol. 18, 246–254 (2016).

    MATH  Google Scholar 

  3. Hofer, M. & Lutolf, M. P. Engineering organoids. Nat. Rev. Mater. 6, 402–420 (2021).

    Google Scholar 

  4. Lancaster, M. A. & Knoblich, J. A. Organogenesis in a dish: modeling development and disease using organoid technologies. Science 345, 1247125 (2014).

    Google Scholar 

  5. Clevers, H. Modeling development and disease with organoids. Cell 165, 1586–1597 (2016).

    MATH  Google Scholar 

  6. Neal, J. T. et al. Organoid modeling of the tumor immune microenvironment. Cell 175, 1972–1988.e16 (2018).

    MATH  Google Scholar 

  7. Sloan, S. A., Andersen, J., Pașca, A. M., Birey, F. & Pașca, S. P. Generation and assembly of human brain region-specific three-dimensional cultures. Nat. Protoc. 13, 2062–2085 (2018).

    MATH  Google Scholar 

  8. Lancaster, M. A. & Knoblich, J. A. Generation of cerebral organoids from human pluripotent stem cells. Nat. Protoc. 9, 2329–2340 (2014).

    MATH  Google Scholar 

  9. Lewis-Israeli, Y. R. et al. Self-assembling human heart organoids for the modeling of cardiac development and congenital heart disease. Nat. Commun. 12, 5142 (2021).

    MATH  Google Scholar 

  10. Guan, Y. et al. Human hepatic organoids for the analysis of human genetic diseases. JCI Insight 2, e94954 (2017).

    MATH  Google Scholar 

  11. Spence, J. R. et al. Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature 470, 105–109 (2011).

    MATH  Google Scholar 

  12. Kruth, J. P., Leu, M. C. & Nakagawa, T. Progress in additive manufacturing and rapid prototyping. CIRP Ann. 47, 525–540 (1998).

    MATH  Google Scholar 

  13. Murphy, S. V. & Atala, A. 3D bioprinting of tissues and organs. Nat. Biotechnol. 32, 773–785 (2014).

    MATH  Google Scholar 

  14. Bernal, P. N. et al. Volumetric bioprinting of complex living-tissue constructs within seconds. Adv. Mater. 31, e1904209 (2019).

    MATH  Google Scholar 

  15. Loterie, D., Delrot, P. & Moser, C. High-resolution tomographic volumetric additive manufacturing. Nat. Commun. 11, 852 (2020).

    Google Scholar 

  16. Kelly, B. E. et al. Volumetric additive manufacturing via tomographic reconstruction. Science 363, 1075–1079 (2019).

    MATH  Google Scholar 

  17. Maharjan, S. et al. Advanced 3D imaging and organoid bioprinting for biomedical research and therapeutic applications. Adv. Drug Deliv. Rev. 208, 115237 (2024).

    Google Scholar 

  18. Hull, S. M., Brunel, L. G. & Heilshorn, S. C. 3D bioprinting of cell-laden hydrogels for improved biological functionality. Adv. Mater. 34, e2103691 (2022).

    MATH  Google Scholar 

  19. Sato, T. et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. Gastroenterology 141, 1762–1772 (2011).

    MATH  Google Scholar 

  20. Takasato, M., Er, P. X., Chiu, H. S. & Little, M. H. Generation of kidney organoids from human pluripotent stem cells. Nat. Protoc. 11, 1681–1692 (2016).

    MATH  Google Scholar 

  21. Sampaziotis, F. et al. Reconstruction of the mouse extrahepatic biliary tree using primary human extrahepatic cholangiocyte organoids. Nat. Med. 23, 954–963 (2017).

    Google Scholar 

  22. Velasco, S. et al. Individual brain organoids reproducibly form cell diversity of the human cerebral cortex. Nature 570, 523–527 (2019).

    MATH  Google Scholar 

  23. Kadoshima, T. et al. Self-organization of axial polarity, inside-out layer pattern, and species-specific progenitor dynamics in human ES cell-derived neocortex. Proc. Natl Acad. Sci. USA 110, 20284–20289 (2013).

    Google Scholar 

  24. Qian, X. et al. Brain-region-specific organoids using mini-bioreactors for modeling ZIKV exposure. Cell 165, 1238–1254 (2016).

    MATH  Google Scholar 

  25. Ranga, A. et al. Neural tube morphogenesis in synthetic 3D microenvironments. Proc. Natl Acad. Sci. USA 113, E6831–E6839 (2016).

    Google Scholar 

  26. Abdel Fattah, A. R. et al. Actuation enhances patterning in human neural tube organoids. Nat. Commun. 12, 3192 (2021).

    MATH  Google Scholar 

  27. Lenne, P. F. et al. Roadmap for the multiscale coupling of biochemical and mechanical signals during development. Phys. Biol. 18, 041501 (2021).

    MATH  Google Scholar 

  28. Aguado, B. A., Mulyasasmita, W., Su, J., Lampe, K. J. & Heilshorn, S. C. Improving viability of stem cells during syringe needle flow through the design of hydrogel cell carriers. Tissue Eng. Pt A 18, 806–815 (2012).

    Google Scholar 

  29. Brassard, J. A., Nikolaev, M., Hübscher, T., Hofer, M. & Lutolf, M. P. Recapitulating macro-scale tissue self-organization through organoid bioprinting. Nat. Mater. 20, 22–29 (2021). This study uses extrusion bioprinting of intestinal stem cells to fabricate intestinal tube-like structures with perfusable lumens.

    Google Scholar 

  30. Lei, M. et al. The mechano-chemical circuit drives skin organoid self-organization. Proc. Natl Acad. Sci. USA 120, e2221982120 (2023).

    Google Scholar 

  31. Ehrig, S. et al. Surface tension determines tissue shape and growth kinetics. Sci. Adv. 5, eaav9394 (2019).

    MATH  Google Scholar 

  32. Riccobelli, D. & Bevilacqua, G. Surface tension controls the onset of gyrification in brain organoids. J. Mech. Phys. Solids 134, 103745 (2020).

    MathSciNet  MATH  Google Scholar 

  33. Fernández, P. A. et al. Surface-tension-induced budding drives alveologenesis in human mammary gland organoids. Nat. Phys. 17, 1130–1136 (2021).

    MATH  Google Scholar 

  34. Roth, J. G. et al. Spatially controlled construction of assembloids using bioprinting. Nat. Commun. 14, 4346 (2023). This study develops a magnetic bioprinting approach to spatially pattern individual, specific neural organoids with minimal structural deformation.

    MATH  Google Scholar 

  35. Bernal, P. N. et al. Volumetric bioprinting of organoids and optically tuned hydrogels to build liver-like metabolic biofactories. Adv. Mater. 34, 2110054 (2022).

    MATH  Google Scholar 

  36. Shin, S. et al. Gelation of uniform interfacial diffusant in embedded 3D printing. Adv. Funct. Mater. 33, 2307435 (2023).

    Google Scholar 

  37. Kratochvil, M. J. et al. Engineered materials for organoid systems. Nat. Rev. Mater. 4, 606–622 (2019).

    MATH  Google Scholar 

  38. Hughes, C. S., Postovit, L. M. & Lajoie, G. A. Matrigel: a complex protein mixture required for optimal growth of cell culture. Proteomics 10, 1886–1890 (2010).

    Google Scholar 

  39. Aisenbrey, E. A. & Murphy, W. L. Synthetic alternatives to Matrigel. Nat. Rev. Mater. 5, 539–551 (2020).

    Google Scholar 

  40. Mulero-Russe, A. & García, A. J. Engineered synthetic matrices for human intestinal organoid culture and therapeutic delivery. Adv. Mater. 36, 2307678 (2024).

    Google Scholar 

  41. Gjorevski, N. et al. Designer matrices for intestinal stem cell and organoid culture. Nature 539, 560–564 (2016).

    Google Scholar 

  42. Hunt, D. R. et al. Engineered matrices enable the culture of human patient-derived intestinal organoids. Adv. Sci. 8, 2004705 (2021).

    Google Scholar 

  43. Cruz-Acuña, R. et al. Synthetic hydrogels for human intestinal organoid generation and colonic wound repair. Nat. Cell Biol. 19, 1326–1335 (2017).

    MATH  Google Scholar 

  44. Nerger, B. A. et al. 3D hydrogel encapsulation regulates nephrogenesis in kidney organoids. Adv. Mater. 36, e2308325 (2024).

    Google Scholar 

  45. Sorrentino, G. et al. Mechano-modulatory synthetic niches for liver organoid derivation. Nat. Commun. 11, 3416 (2020).

    MATH  Google Scholar 

  46. Hernandez-Gordillo, V. et al. Fully synthetic matrices for in vitro culture of primary human intestinal enteroids and endometrial organoids. Biomaterials 254, 120125 (2020).

    Google Scholar 

  47. Suhar, R. A., Huang, M. S., Navarro, R. S., Aviles Rodriguez, G. & Heilshorn, S. C. A library of elastin-like proteins with tunable matrix ligands for in vitro 3D neural cell culture. Biomacromolecules 24, 5926–5939 (2023).

    Google Scholar 

  48. LeSavage, B. L. et al. Engineered matrices reveal stiffness-mediated chemoresistance in patient-derived pancreatic cancer organoids. Nat. Mater. 23, 1138–1149 (2024).

    MATH  Google Scholar 

  49. Ligorio, C. & Mata, A. Synthetic extracellular matrices with function-encoding peptides. Nat. Rev. Bioeng. 1, 518–536 (2023).

    Google Scholar 

  50. Roth, J. G. et al. Advancing models of neural development with biomaterials. Nat. Rev. Neurosci. 22, 593–615 (2021).

    MATH  Google Scholar 

  51. Chaudhuri, O., Cooper-White, J., Janmey, P. A., Mooney, D. J. & Shenoy, V. B. Effects of extracellular matrix viscoelasticity on cellular behaviour. Nature 584, 535–546 (2020).

    Google Scholar 

  52. Elosegui-Artola, A. et al. Matrix viscoelasticity controls spatiotemporal tissue organization. Nat. Mater. 22, 117–127 (2023).

    Google Scholar 

  53. Peng, Y. H. et al. Dynamic matrices with DNA-encoded viscoelasticity for cell and organoid culture. Nat. Nanotechnol. 18, 1463–1473 (2023).

    MATH  Google Scholar 

  54. Yavitt, F. M. et al. In situ modulation of intestinal organoid epithelial curvature through photoinduced viscoelasticity directs crypt morphogenesis. Sci. Adv. 9, eadd5668 (2023).

    Google Scholar 

  55. Haw, M. D. Jamming, two-fluid behavior, and ‘self-filtration’ in concentrated particulate suspensions. Phys. Rev. Lett. 92, 185506 (2004).

    MATH  Google Scholar 

  56. Ho, D. L. L. et al. Large-scale production of wholly cellular bioinks via the optimization of human induced pluripotent stem cell aggregate culture in automated bioreactors. Adv. Healthc. Mater. 11, e2201138 (2022).

    Google Scholar 

  57. Skylar-Scott, M. A. et al. Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels. Sci. Adv. 5, eaaw2459 (2019). This study incorporates spheroids and organoids as the support bath, into which a sacrificial ink can be printed to yield vascular channels embedded in a high cell density fused tissue-like construct.

    Google Scholar 

  58. Steinberg, M. S. Differential adhesion in morphogenesis: a modern view. Curr. Opin. Genet. Dev. 17, 281–286 (2007).

    MATH  Google Scholar 

  59. Mironov, V. et al. Organ printing: tissue spheroids as building blocks. Biomaterials 30, 2164–2174 (2009).

    MATH  Google Scholar 

  60. Kelley, K. W. & Pașca, S. P. Human brain organogenesis: toward a cellular understanding of development and disease. Cell 185, 42–61 (2022).

    MATH  Google Scholar 

  61. Pașca, S. P. et al. A nomenclature consensus for nervous system organoids and assembloids. Nature 609, 907–910 (2022).

    MATH  Google Scholar 

  62. Bagley, J. A., Reumann, D., Bian, S., Lévi-Strauss, J. & Knoblich, J. A. Fused cerebral organoids model interactions between brain regions. Nat. Methods 14, 743–751 (2017).

    Google Scholar 

  63. Birey, F. et al. Assembly of functionally integrated human forebrain spheroids. Nature 545, 54–59 (2017).

    Google Scholar 

  64. Xiang, Y. et al. Fusion of regionally specified hPSC-derived organoids models human brain development and interneuron migration. Cell Stem Cell 21, 383–398.e7 (2017).

    MATH  Google Scholar 

  65. Sinha, S. et al. Laminin-associated integrins mediate diffuse intrinsic pontine glioma infiltration and therapy response within a neural assembloid model. Acta Neuropathol. Commun. 12, 71 (2024).

    MATH  Google Scholar 

  66. Sachs, N., Tsukamoto, Y., Kujala, P., Peters, P. J. & Clevers, H. Intestinal epithelial organoids fuse to form self-organizing tubes in floating collagen gels. Development 144, 1107–1112 (2017).

    Google Scholar 

  67. Jakab, K. et al. Relating cell and tissue mechanics: implications and applications. Dev. Dyn. 237, 2438–2449 (2008).

    MATH  Google Scholar 

  68. Fleming, P. A. et al. Fusion of uniluminal vascular spheroids: a model for assembly of blood vessels. Dev. Dyn. 239, 398–406 (2010).

    MATH  Google Scholar 

  69. Frostad, J. M., Collins, M. C. & Leal, L. G. Cantilevered-capillary force apparatus for measuring multiphase fluid interactions. Langmuir 29, 4715–4725 (2013).

    Google Scholar 

  70. Chatzigiannakis, E., Chen, Y., Bachnak, R., Dutcher, C. S. & Vermant, J. Studying coalescence at different lengthscales: from films to droplets. Rheol. Acta 61, 745–759 (2022).

    Google Scholar 

  71. Handorf, A. M., Zhou, Y., Halanski, M. A. & Li, W. J. Tissue stiffness dictates development, homeostasis, and disease progression. Organogenesis 11, 1–15 (2015).

    Google Scholar 

  72. Wolf, K. J., Weiss, J. D., Uzel, S. G. M., Skylar-Scott, M. A. & Lewis, J. A. Biomanufacturing human tissues via organ building blocks. Cell Stem Cell 29, 667–677 (2022).

    Google Scholar 

  73. Kupfer, M. E. et al. In situ expansion, differentiation, and electromechanical coupling of human cardiac muscle in a 3D bioprinted, chambered organoid. Circ. Res. 127, 207–224 (2020).

    MATH  Google Scholar 

  74. Lawlor, K. T. et al. Cellular extrusion bioprinting improves kidney organoid reproducibility and conformation. Nat. Mater. 20, 260–271 (2021). This study uses extrusion bioprinting for the reproducible and scalable generation of kidney organoids.

    MATH  Google Scholar 

  75. Groll, J. et al. Biofabrication: reappraising the definition of an evolving field. Biofabrication 8, 013001 (2016).

    MATH  Google Scholar 

  76. Jakab, K. et al. Tissue engineering by self-assembly of cells printed into topologically defined structures. Tissue Eng. Part A 14, 413–421 (2008). This study uses capillary extrusion of pre-formed spheroids to fabricate fused spheroid assemblies of defined geometries.

    MATH  Google Scholar 

  77. Norotte, C., Marga, F. S., Niklason, L. E. & Forgacs, G. Scaffold-free vascular tissue engineering using bioprinting. Biomaterials 30, 5910–5917 (2009).

    Google Scholar 

  78. Mekhileri, N. V. et al. Automated 3D bioassembly of micro-tissues for biofabrication of hybrid tissue engineered constructs. Biofabrication 10, 024103 (2018).

    MATH  Google Scholar 

  79. Mekhileri, N. V. et al. Biofabrication of modular spheroids as tumor-scale microenvironments for drug screening. Adv. Healthc. Mater. 12, e2201581 (2023).

    Google Scholar 

  80. Goulart, E. et al. 3D bioprinting of liver spheroids derived from human induced pluripotent stem cells sustain liver function and viability in vitro. Biofabrication 12, 015010 (2019).

    MATH  Google Scholar 

  81. De Moor, L. et al. Engineering microvasculature by 3D bioprinting of prevascularized spheroids in photo-crosslinkable gelatin. Biofabrication 13, 045021 (2021).

    MATH  Google Scholar 

  82. LeSavage, B. L., Suhar, R. A., Broguiere, N., Lutolf, M. P. & Heilshorn, S. C. Next-generation cancer organoids. Nat. Mater. 21, 143–159 (2022).

    Google Scholar 

  83. Cruz-Acuña, R. et al. Engineered hydrogel reveals contribution of matrix mechanics to esophageal adenocarcinoma and identifies matrix-activated therapeutic targets. J. Clin. Invest. 133, e168146 (2023).

    Google Scholar 

  84. Swaminathan, S., Hamid, Q., Sun, W. & Clyne, A. M. Bioprinting of 3D breast epithelial spheroids for human cancer models. Biofabrication 11, 025003 (2019).

    Google Scholar 

  85. Kim, E. et al. Creation of bladder assembloids mimicking tissue regeneration and cancer. Nature 588, 664–669 (2020).

    MATH  Google Scholar 

  86. Andersen, J. et al. Generation of functional human 3D cortico-motor assembloids. Cell 183, 1913–1929.e26 (2020).

    MATH  Google Scholar 

  87. Miura, Y. et al. Generation of human striatal organoids and cortico-striatal assembloids from human pluripotent stem cells. Nat. Biotechnol. 38, 1421–1430 (2020).

    MATH  Google Scholar 

  88. Schmidt, C. et al. Multi-chamber cardioids unravel human heart development and cardiac defects. Cell 186, 5587–5605.e27 (2023).

    MATH  Google Scholar 

  89. Kim, J.-I. et al. Human assembloid model of the ascending neural sensory pathway. Preprint at bioRxiv https://doi.org/10.1101/2024.03.11.584539 (2024).

  90. Nakayama, K. in Biofabrication (eds G. Forgacs & W. Sun) 1–21 (William Andrew Publishing, 2013).

  91. Itoh, M. et al. Scaffold-free tubular tissues created by a bio-3D printer undergo remodeling and endothelialization when implanted in rat aortae. PLoS ONE 10, e0136681 (2015). This study introduces the Kenzan method, which is a method of spheroid assembly that involves skewering pre-formed spheroids on a microneedle array.

    Google Scholar 

  92. Yamasaki, A. et al. Osteochondral regeneration using constructs of mesenchymal stem cells made by bio three-dimensional printing in mini-pigs. J. Orthop. Res. 37, 1398–1408 (2019).

    MATH  Google Scholar 

  93. Imamura, T. et al. Biofabricated structures reconstruct functional urinary bladders in radiation-injured rat bladders. Tissue Eng. Part A 24, 1574–1587 (2018).

    MATH  Google Scholar 

  94. Murata, D., Arai, K. & Nakayama, K. Scaffold-free bio-3D printing using spheroids as ‘Bio-Inks’ for tissue (re-)construction and drug response tests. Adv. Healthc. Mater. 9, 1901831 (2020).

    Google Scholar 

  95. Ayan, B. et al. Aspiration-assisted bioprinting for precise positioning of biologics. Sci. Adv. 6, eaaw5111 (2020). This study describes aspiration-assisted bioprinting for spatially positioning spheroids by using vacuum aspiration to select and deposit individual spheroids.

    MATH  Google Scholar 

  96. Heo, D. N. et al. Aspiration-assisted bioprinting of co-cultured osteogenic spheroids for bone tissue engineering. Biofabrication 13, 015013 (2020).

    MATH  Google Scholar 

  97. Daly, A. C., Davidson, M. D. & Burdick, J. A. 3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels. Nat. Commun. 12, 753 (2021).

    Google Scholar 

  98. Kim, W., Gwon, Y., Park, S., Kim, H. & Kim, J. Therapeutic strategies of three-dimensional stem cell spheroids and organoids for tissue repair and regeneration. Bioact. Mater. 19, 50–74 (2023).

    MATH  Google Scholar 

  99. Brunel, L. G., Hull, S. M. & Heilshorn, S. C. Engineered assistive materials for 3D bioprinting: support baths and sacrificial inks. Biofabrication 14, 032001 (2022).

    Google Scholar 

  100. Becker, M., Gurian, M., Schot, M. & Leijten, J. Aqueous two-phase enabled low viscosity 3D (LoV3D) bioprinting of living matter. Adv. Sci. 10, e2204609 (2023).

    Google Scholar 

  101. Sakai, S. & Morita, T. One-step FRESH bioprinting of low-viscosity silk fibroin inks. ACS Biomater. Sci. Eng. 8, 2589–2597 (2022).

    MATH  Google Scholar 

  102. Tan, E. Y. S., Suntornnond, R. & Yeong, W. Y. High-resolution novel indirect bioprinting of low-viscosity cell-laden hydrogels via model-support bioink interaction. 3D Print. Addit. Manuf. 8, 69–78 (2021).

    Google Scholar 

  103. Brunel, L. G. et al. Embedded 3D bioprinting of collagen inks into microgel baths to control hydrogel microstructure and cell spreading. Adv. Healthc. Mater. 13, 2303325 (2024).

    MATH  Google Scholar 

  104. Jakab, K., Damon, B., Neagu, A., Kachurin, A. & Forgacs, G. Three-dimensional tissue constructs built by bioprinting. Biorheology 43, 509–513 (2006).

    MATH  Google Scholar 

  105. Wu, W., DeConinck, A. & Lewis, J. A. Omnidirectional printing of 3D microvascular networks. Adv. Mater. 23, H178–H183 (2011).

    MATH  Google Scholar 

  106. Bhattacharjee, T. et al. Writing in the granular gel medium. Sci. Adv. 1, e1500655 (2015).

    MATH  Google Scholar 

  107. Highley, C. B., Rodell, C. B. & Burdick, J. A. Direct 3D printing of shear-thinning hydrogels into self-healing hydrogels. Adv. Mater. 27, 5075–5079 (2015).

    Google Scholar 

  108. Hinton, T. J. et al. Three-dimensional printing of complex biological structures by freeform reversible embedding of suspended hydrogels. Sci. Adv. 1, e1500758 (2015).

    MATH  Google Scholar 

  109. Lee, A. et al. 3D bioprinting of collagen to rebuild components of the human heart. Science 365, 482–487 (2019).

    MATH  Google Scholar 

  110. Noor, N. et al. 3D printing of personalized thick and perfusable cardiac patches and hearts. Adv. Sci. 6, 1900344 (2019).

    Google Scholar 

  111. Moxon, S. R. et al. Suspended manufacture of biological structures. Adv. Mater. 29, 1605594 (2017).

    MATH  Google Scholar 

  112. Compaan, A. M., Song, K. & Huang, Y. Gellan fluid gel as a versatile support bath material for fluid extrusion bioprinting. ACS Appl. Mater. Interfaces 11, 5714–5726 (2019).

    MATH  Google Scholar 

  113. Jin, Y., Compaan, A., Chai, W. & Huang, Y. Functional nanoclay suspension for printing-then-solidification of liquid materials. ACS Appl. Mater. Interfaces 9, 20057–20066 (2017).

    MATH  Google Scholar 

  114. Morley, C. D. et al. Quantitative characterization of 3D bioprinted structural elements under cell generated forces. Nat. Commun. 10, 3029 (2019).

    MATH  Google Scholar 

  115. Shin, S. & Hyun, J. Matrix-assisted three-dimensional printing of cellulose nanofibers for paper microfluidics. ACS Appl. Mater. Interfaces 9, 26438–26446 (2017).

    MATH  Google Scholar 

  116. Bertassoni, L. E. et al. Direct-write bioprinting of cell-laden methacrylated gelatin hydrogels. Biofabrication 6, 024105 (2014).

    MATH  Google Scholar 

  117. Bertassoni, L. E. et al. Hydrogel bioprinted microchannel networks for vascularization of tissue engineering constructs. Lab Chip 14, 2202–2211 (2014).

    MATH  Google Scholar 

  118. Hammer, J., Han, L. H., Tong, X. & Yang, F. A facile method to fabricate hydrogels with microchannel-like porosity for tissue engineering. Tissue Eng. Part C Methods 20, 169–176 (2014).

    Google Scholar 

  119. Wang, X. Y. et al. Engineering interconnected 3D vascular networks in hydrogels using molded sodium alginate lattice as the sacrificial template. Lab Chip 14, 2709–2716 (2014).

    MATH  Google Scholar 

  120. Visser, J. et al. Biofabrication of multi-material anatomically shaped tissue constructs. Biofabrication 5, 035007 (2013).

    MATH  Google Scholar 

  121. Pimentel, C. R. et al. Three-dimensional fabrication of thick and densely populated soft constructs with complex and actively perfused channel network. Acta Biomater. 65, 174–184 (2018).

    MATH  Google Scholar 

  122. Miller, J. S. et al. Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues. Nat. Mater. 11, 768–774 (2012).

    MATH  Google Scholar 

  123. Ozbolat, V., Dey, M., Ayan, B. & Ozbolat, I. T. Extrusion-based printing of sacrificial Carbopol ink for fabrication of microfluidic devices. Biofabrication 11, 034101 (2019).

    Google Scholar 

  124. Seymour, A. J., Shin, S. & Heilshorn, S. C. 3D printing of microgel scaffolds with tunable void fraction to promote cell infiltration. Adv. Healthc. Mater. 10, e2100644 (2021).

    Google Scholar 

  125. Ying, G. L. et al. Aqueous two-phase emulsion bioink-enabled 3D bioprinting of porous hydrogels. Adv. Mater. 30, e1805460 (2018).

    Google Scholar 

  126. Hull, S. M. et al. 3D bioprinting using UNIversal Orthogonal Network (UNION) Bioinks. Adv. Funct. Mater. 31, 2007983 (2021).

    MathSciNet  Google Scholar 

  127. Jeon, O. et al. Individual cell-only bioink and photocurable supporting medium for 3D printing and generation of engineered tissues with complex geometries. Mater. Horiz. 6, 1625–1631 (2019).

    MATH  Google Scholar 

  128. Gao, T. & Hu, H. H. Deformation of elastic particles in viscous shear flow. J. Comput. Phys. 228, 2132–2151 (2009).

    MATH  Google Scholar 

  129. Chen, Y.-L. Inertia- and deformation-driven migration of a soft particle in confined shear and Poiseuille flow. RSC Adv. 4, 17908–17916 (2014).

    MATH  Google Scholar 

  130. Dubbin, K., Hori, Y., Lewis, K. K. & Heilshorn, S. C. Dual-stage crosslinking of a gel-phase bioink improves cell viability and homogeneity for 3D bioprinting. Adv. Healthc. Mater. 5, 2488–2492 (2016).

    Google Scholar 

  131. Flégeau, K., Puiggali-Jou, A. & Zenobi-Wong, M. Cartilage tissue engineering by extrusion bioprinting utilizing porous hyaluronic acid microgel bioinks. Biofabrication 14, 034105 (2022).

    Google Scholar 

  132. Park, J. Y. et al. Development of a functional airway-on-a-chip by 3D cell printing. Biofabrication 11, 015002 (2018).

    MATH  Google Scholar 

  133. Kolesky, D. B., Homan, K. A., Skylar-Scott, M. A. & Lewis, J. A. Three-dimensional bioprinting of thick vascularized tissues. Proc. Natl Acad. Sci. USA 113, 3179–3184 (2016).

    Google Scholar 

  134. Gao, G., Park, J. Y., Kim, B. S., Jang, J. & Cho, D. W. Coaxial cell printing of freestanding, perfusable, and functional in vitro vascular models for recapitulation of native vascular endothelium pathophysiology. Adv. Healthc. Mater. 7, e1801102 (2018).

    Google Scholar 

  135. Hunziker, E. B., Quinn, T. M. & Häuselmann, H. J. Quantitative structural organization of normal adult human articular cartilage. Osteoarthr. Cartil. 10, 564–572 (2002).

    Google Scholar 

  136. Quinn, T. M., Häuselmann, H. J., Shintani, N. & Hunziker, E. B. Cell and matrix morphology in articular cartilage from adult human knee and ankle joints suggests depth-associated adaptations to biomechanical and anatomical roles. Osteoarthr. Cartil. 21, 1904–1912 (2013).

    Google Scholar 

  137. Blouin, A., Bolender, R. P. & Weibel, E. R. Distribution of organelles and membranes between hepatocytes and nonhepatocytes in the rat liver parenchyma. A stereological study. J. Cell Biol. 72, 441–455 (1977).

    Google Scholar 

  138. Gullino, P. M., Grantham, F. H. & Smith, S. H. The interstitial water space of tumors. Cancer Res. 25, 727–731 (1965).

    MATH  Google Scholar 

  139. Donahue, K. M. et al. Dynamic Gd-DTPA enhanced MRI measurement of tissue cell volume fraction. Magn. Reson. Med. 34, 423–432 (1995).

    MATH  Google Scholar 

  140. Thulborn, K. et al. Quantitative sodium MRI of the human brain at 9.4 T provides assessment of tissue sodium concentration and cell volume fraction during normal aging. NMR Biomed. 29, 137–143 (2016).

    Google Scholar 

  141. Thulborn, K. R. Gender differences in cell volume fraction (CVF): a structural parameter reflecting the energy efficiency of maintaining the resting membrane potential. NMR Biomed. 35, e4693 (2022).

    Google Scholar 

  142. Stankey, P. P. et al. Embedding biomimetic vascular networks via coaxial sacrificial writing into functional tissue. Adv. Mater. 36, e2401528 (2024).

    Google Scholar 

  143. Skylar-Scott, M. A. et al. Orthogonally induced differentiation of stem cells for the programmatic patterning of vascularized organoids and bioprinted tissues. Nat. Biomed. Eng. 6, 449–462 (2022).

    Google Scholar 

  144. Ritzau-Reid, K. I. et al. Microfibrous scaffolds guide stem cell lumenogenesis and brain organoid engineering. Adv. Mater. 35, e2300305 (2023).

    Google Scholar 

  145. Trentesaux, C., Yamada, T., Klein, O. D. & Lim, W. A. Harnessing synthetic biology to engineer organoids and tissues. Cell Stem Cell 30, 10–19 (2023).

    Google Scholar 

  146. Cederquist, G. Y. et al. Specification of positional identity in forebrain organoids. Nat. Biotechnol. 37, 436–444 (2019).

    MATH  Google Scholar 

  147. Toda, S., Blauch, L. R., Tang, S. K. Y., Morsut, L. & Lim, W. A. Programming self-organizing multicellular structures with synthetic cell–cell signaling. Science 361, 156–162 (2018).

    Google Scholar 

  148. Bao, M. et al. Stem cell-derived synthetic embryos self-assemble by exploiting cadherin codes and cortical tension. Nat. Cell Biol. 24, 1341–1349 (2022).

    MATH  Google Scholar 

  149. Cakir, B. et al. Engineering of human brain organoids with a functional vascular-like system. Nat. Methods 16, 1169–1175 (2019).

    MATH  Google Scholar 

  150. Ribezzi, D. et al. Shaping synthetic multicellular and complex multimaterial tissues via embedded extrusion-volumetric printing of microgels. Adv. Mater. 35, e2301673 (2023).

    MATH  Google Scholar 

  151. Tringides, C. M. et al. Tunable conductive hydrogel scaffolds for neural cell differentiation. Adv. Healthc. Mater. 12, 2202221 (2023).

    Google Scholar 

  152. Paunović, N. et al. 4D printing of biodegradable elastomers with tailorable thermal response at physiological temperature. J. Control. Release 361, 417–426 (2023).

    MATH  Google Scholar 

  153. Wirtz, D. Particle-tracking microrheology of living cells: principles and applications. Annu. Rev. Biophys. 38, 301–326 (2009).

    MATH  Google Scholar 

  154. Squires, T. M. & Mason, T. G. Fluid mechanics of microrheology. Annu. Rev. Fluid Mech. 42, 413–438 (2010).

    MATH  Google Scholar 

  155. McGlynn, J. A., Wu, N. & Schultz, K. M. Multiple particle tracking microrheological characterization: fundamentals, emerging techniques and applications. J. Appl. Phys. 127, 201101 (2020).

    Google Scholar 

  156. Schultz, K. M. & Furst, E. M. Microrheology of biomaterial hydrogelators. Soft Matter 8, 6198–6205 (2012).

    MATH  Google Scholar 

  157. Woolley, L., Burbidge, A., Vermant, J. & Christakopoulos, F. A microrheological examination of insulin-secreting β-cells in healthy and diabetic-like conditions. Soft Matter 20, 3464–3472 (2024).

    Google Scholar 

  158. Mason, T. G. Estimating the viscoelastic moduli of complex fluids using the generalized Stokes–Einstein equation. Rheol. Acta 39, 371–378 (2000).

    MATH  Google Scholar 

  159. Guo, M. et al. Probing the stochastic, motor-driven properties of the cytoplasm using force spectrum microscopy. Cell 158, 822–832 (2014).

    MATH  Google Scholar 

  160. Sriram, I., Furst, E. M., DePuit, R. J. & Squires, T. M. Small amplitude active oscillatory microrheology of a colloidal suspension. J. Rheol. 53, 357–381 (2009).

    MATH  Google Scholar 

  161. Efremov, Y. M., Okajima, T. & Raman, A. Measuring viscoelasticity of soft biological samples using atomic force microscopy. Soft Matter 16, 64–81 (2020).

    MATH  Google Scholar 

  162. Mattice, J. M., Lau, A. G., Oyen, M. L. & Kent, R. W. Spherical indentation load-relaxation of soft biological tissues. J. Mater. Res. 21, 2003–2010 (2006).

    Google Scholar 

  163. Alcaraz, J. et al. Laminin and biomimetic extracellular elasticity enhance functional differentiation in mammary epithelia. EMBO J. 27, 2829–2838 (2008).

    MATH  Google Scholar 

  164. Lyu, Q. et al. A soft and ultrasensitive force sensing diaphragm for probing cardiac organoids instantaneously and wirelessly. Nat. Commun. 13, 7259 (2022).

    MATH  Google Scholar 

  165. Vyas, V., Solomon, M., D’Souza, G. G. M. & Huey, B. D. Nanomechanical analysis of extracellular matrix and cells in multicellular spheroids. Cell Mol. Bioeng. 12, 203–214 (2019).

    Google Scholar 

  166. Kwon, E.-Y., Kim, Y.-T. & Kim, D.-E. Investigation of penetration force of living cell using an atomic force microscope. J. Mech. Sci. Technol. 23, 1932–1938 (2009).

    MATH  Google Scholar 

  167. Hochmuth, R. M. Micropipette aspiration of living cells. J. Biomech. 33, 15–22 (2000).

    MATH  Google Scholar 

  168. Barnes, H. A. Rheology: principles, measurements and applications. Powder Technol. 86, 313 (1996).

    MATH  Google Scholar 

  169. Ewoldt, R. H., Johnston, M. T. & Caretta, L. M. in Complex Fluids in Biological Systems: Experiment, Theory, and Computation (ed. Spagnolie, S. E.) 207–241 (Springer New York, 2015).

  170. Du, J. et al. A visual, in-expensive, and wireless capillary rheometer for characterizing wholly-cellular bioinks. Small 20, e2304778 (2024).

    Google Scholar 

  171. Tomasina, C., Bodet, T., Mota, C., Moroni, L. & Camarero-Espinosa, S. Bioprinting vasculature: materials, cells and emergent techniques. Materials 12, 2701 (2019).

    Google Scholar 

  172. Chen, E. P., Toksoy, Z., Davis, B. A. & Geibel, J. P. 3D bioprinting of vascularized tissues for in vitro and in vivo applications. Front. Bioeng. Biotechnol. 9, 664188 (2021).

    MATH  Google Scholar 

  173. Carmeliet, P. & Jain, R. K. Angiogenesis in cancer and other diseases. Nature 407, 249–257 (2000).

    MATH  Google Scholar 

  174. Jain, R. K., Au, P., Tam, J., Duda, D. G. & Fukumura, D. Engineering vascularized tissue. Nat. Biotechnol. 23, 821–823 (2005).

    Google Scholar 

  175. Datta, P., Ayan, B. & Ozbolat, I. T. Bioprinting for vascular and vascularized tissue biofabrication. Acta Biomater. 51, 1–20 (2017).

    MATH  Google Scholar 

  176. Zhang, Y., Yu, Y., Chen, H. & Ozbolat, I. T. Characterization of printable cellular micro-fluidic channels for tissue engineering. Biofabrication 5, 025004 (2013).

    Google Scholar 

  177. Miri, A. K. et al. Multiscale bioprinting of vascularized models. Biomaterials 198, 204–216 (2019).

    MATH  Google Scholar 

  178. Zhang, Y. et al. Recent advances in 3D bioprinting of vascularized tissues. Mater. Des. 199, 109398 (2021).

    MATH  Google Scholar 

  179. Wei, Z. et al. Hydrogels with tunable mechanical plasticity regulate endothelial cell outgrowth in vasculogenesis and angiogenesis. Nat. Commun. 14, 8307 (2023).

    MATH  Google Scholar 

  180. Luttun, A. et al. Revascularization of ischemic tissues by PlGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Flt1. Nat. Med. 8, 831–840 (2002).

    Google Scholar 

  181. Richardson, T. P., Peters, M. C., Ennett, A. B. & Mooney, D. J. Polymeric system for dual growth factor delivery. Nat. Biotechnol. 19, 1029–1034 (2001).

    Google Scholar 

  182. Jain, R. K. Molecular regulation of vessel maturation. Nat. Med. 9, 685–693 (2003).

    MATH  Google Scholar 

  183. Macklin, B. L. et al. Intrinsic epigenetic control of angiogenesis in induced pluripotent stem cell-derived endothelium regulates vascular regeneration. npj Regen. Med. 7, 28 (2022).

    MATH  Google Scholar 

  184. Inglis, S., Kanczler, J. M. & Oreffo, R. O. C. 3D human bone marrow stromal and endothelial cell spheres promote bone healing in an osteogenic niche. FASEB J. 33, 3279–3290 (2019).

    Google Scholar 

  185. Salmon, I. et al. Engineering neurovascular organoids with 3D printed microfluidic chips. Lab Chip 22, 1615–1629 (2022).

    MATH  Google Scholar 

  186. Stresser, D. M. et al. Towards in vitro models for reducing or replacing the use of animals in drug testing. Nat. Biomed. Eng. 8, 930–935 (2023).

    MATH  Google Scholar 

  187. Mariani, J. et al. FOXG1-dependent dysregulation of GABA/glutamate neuron differentiation in autism spectrum disorders. Cell 162, 375–390 (2015).

    MATH  Google Scholar 

  188. Vlachogiannis, G. et al. Patient-derived organoids model treatment response of metastatic gastrointestinal cancers. Science 359, 920–926 (2018).

    MATH  Google Scholar 

  189. Sachs, N. et al. A living biobank of breast cancer organoids captures disease heterogeneity. Cell 172, 373–386.e10 (2018).

    MATH  Google Scholar 

  190. LeBlanc, V. G. et al. Single-cell landscapes of primary glioblastomas and matched explants and cell lines show variable retention of inter- and intratumor heterogeneity. Cancer Cell 40, 379–392.e9 (2022).

    MATH  Google Scholar 

  191. Co, J. Y., Klein, J. A., Kang, S. & Homan, K. A. Toward inclusivity in preclinical drug development: a proposition to start with intestinal organoids. Adv. Biol. 7, e2200333 (2023).

    Google Scholar 

  192. Vandana, J. J., Manrique, C., Lacko, L. A. & Chen, S. Human pluripotent-stem-cell-derived organoids for drug discovery and evaluation. Cell Stem Cell 30, 571–591 (2023).

    MATH  Google Scholar 

  193. Chen, X. et al. Antisense oligonucleotide therapeutic approach for Timothy syndrome. Nature 628, 818–825 (2024).

    MATH  Google Scholar 

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Acknowledgements

The authors thank S. Pasca for helpful conversations about organoid biology and editing of the manuscript. This work was funded in part by a Stanford Cancer Institute Innovation Seed Grant, a Stanford Maternal and Child Health Research Institute Uytengsu-Hamilton 22q11 Neuropsychiatry Seed Grant, the National Institutes of Health (NIH) (R01 HL142718, R01 DK129309 and R01 EB027171), the Advanced Research Projects Agency for Health (ARPA-H) under award number AY1AX000002 and the National Science Foundation (NSF) (DMR 2103812, CBET 2033302). M.S.H. was supported by a NIH F31 Pre-doctoral Fellowship (NS132505) and a Sarafan ChEM-H O’Leary-Thiry Fellowship. F.C. was supported by a Swiss National Science Foundation Postdoc Mobility Fellowship (PN210723).

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  1. These authors contributed equally: Michelle S. Huang, Fotis Christakopoulos.

Authors and Affiliations

  1. Department of Chemical Engineering, Stanford University, Stanford, CA, USA

    Michelle S. Huang

  2. The Institute for Chemistry, Engineering & Medicine for Human Health (Sarafan ChEM-H), Stanford University, Stanford, CA, USA

    Michelle S. Huang & Sarah C. Heilshorn

  3. Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA

    Fotis Christakopoulos & Sarah C. Heilshorn

  4. Complex In Vitro Systems, Safety Assessment, Genentech Inc., South San Francisco, CA, USA

    Julien G. Roth

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M.S.H., F.C. and S.C.H. conceptualized the review. M.S.H., F.C. and J.G.R. researched data for the article and contributed to discussion of content and writing. All authors reviewed and edited the manuscript before submission.

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Correspondence to Sarah C. Heilshorn.

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S.C.H. and J.G.R. are listed as inventors on a patent application (US 2023-0357685 A1) that covers some of the technologies described in this manuscript. The other authors declare no competing interests.

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Huang, M.S., Christakopoulos, F., Roth, J.G. et al. Organoid bioprinting: from cells to functional tissues. Nat Rev Bioeng 3, 126–142 (2025). https://doi.org/10.1038/s44222-024-00268-0

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