Toward system-level integration of organoids for regenerative medicine

Abstract

Regenerating complex human tissues requires proper cellular assembly and the recapitulation of coordinated functions across multiple biological levels. Organoids, as self-organized three-dimensional cellular structures, provide powerful models for rebuilding organ architecture and studying developmental processes. However, their regenerative potential remains limited by the lack of vascular, neural, and immune integration, which are essential for tissue development, homeostasis, and repair. Recent studies indicate that the progression from tissue-level organization to organ-level coordination and ultimately to system-level functionality depends on dynamic intercellular communication, feedback signaling, and niche interactions. The integration of biochemical, biomechanical, and bioelectrical cues enables multicellular systems to achieve synchronized growth, patterning, and functional adaptation. Complementary bioengineering strategies further guide these intrinsic processes by modulating spatial organization, microenvironmental signals, and intercellular connectivity. This review summarizes emerging methodologies and molecular mechanisms underlying system-level integration in organoids and discusses how these biological processes may bridge the gap between in vitro morphogenesis and in vivo functional regeneration.

Keywords

Organoids / Integrated organoids / Regenerative medicine / Tissue repair / Vascularized / Inner vated / Immune-competent

References

[1] Marsee A, Roos FJM, Verstegen MMA, Marsee A, Roos F, Verstegen M, et al. Building consensus on definition and nomenclature of hepatic, pancreatic, and biliary organoids. Cell Stem Cell 2021; 28: 816-32. https://doi.org/10.1016/j.stem.2021.04.005.

[2] Boers SN, Bredenoord AL. Consent for governance in the ethical use of organoids. Nat Cell Biol 2018; 20: 642-5. https://doi.org/10.1038/s41556-018-0112-5.

[3] Zhao Z, Chen X, Dowbaj AM, Sljukic A, Bratlie K, Lin L, et al. Organoids. Nature reviews methods primers. Springer Nature 2022; 2: 94. https://doi.org/10.1038/s43586-022-00174-y.

[4] Bai L, Jing Y, Reis RL, Chen X, Su J, Liu C. Organoid research: theory, technology, and therapeutics. Organoid Research 2025; 1: 025040007. https://doi.org/10.36922/OR025040007.

[5] Bai L, Reis RL, Chen X, Su J, Liu C. Organoid research: advanced models, precision medicine, and translational medicine. Organoid Research 2025; 1: 025060009. https://doi.org/10.36922/OR025060009.

[6] Hosseini M, Koehler KR, Shafiee A. Biofabrication of human skin with its appendages. Adv Healthc Mater 2022; 11: e2201626. https://doi.org/10.1002/adhm.202201626.

[7] Kang S, Kim D, Lee J, Takayama S, Park JY. Engineered microsystems for spheroid and organoid studies. Adv Healthc Mater 2021; 10: e2001284. https://doi.org/10.1002/adhm.202001284.

[8] Deng S, Zhu F, Dai K, Wang J, Liu C. Harvest of functional mesenchymal stem cells derived from in vivo osteo-organoids. Biomater Transl 2023; 4: 270-9. https://doi.org/10.12336/biomatertransl.2023.04.006.

[9] Lei M, Harn HI-C, Li Q, Jiang J, Wu W, Zhou W, et al. The mechano-chemical circuit drives skin organoid self-organization. Proc Natl Acad Sci 2023; 120: e2221982120. https://doi.org/10.1073/pnas.2221982120.

[10] Li T, Li X, Xiang X, Huang J, Shen X, Wang M, et al. Regenerative hair pigmentation via skin organoids: adaptive patterning mediated by collagen VI and semaphorin 3C. Adv Sci 2025; 12: e02436. https://doi.org/10.1002/advs.202502436.

[11] Tang X-Y, Wu S, Wang D, Chu C, Hong Y, Tao M, et al. Human organoids in basic research and clinical applications. Signal Transduct Target Ther 2022; 7: 168. https://doi.org/10.1038/s41392-022-01024-9.

[12] Verstegen MMA, Coppes RP, Beghin A, De Coppi P, Gerli MFM, de Graeff N, et al. Clinical applications of human organoids. Nat Med 2025; 31: 409-21. https://doi.org/10.1038/s41591-024-03489-3.

[13] Revah O, Gore F, Kelley KW, Andersen J, Sakai N, Chen X, et al. Maturation and circuit integration of transplanted human cortical organoids. Nature 2022; 610: 319-26. https://doi.org/10.1038/s41586-022-05277-w.

[14] Zhou Z, Liu J, Xiong T, Liu Y, Tuan RS, Li ZA. Engineering innervated musculoskeletal tissues for regenerative orthopedics and disease modeling. Small 2024; 20: e2310614. https://doi.org/10.1002/smll.202310614.

[15] Recaldin T, Steinacher L, Gjeta B, Harter MF, Adam L, Kromer K, et al. Human organoids with an autologous tissue-resident immune compartment. Nature 2024; 633: 165-73. https://doi.org/10.1038/s41586-024-07791-5.

[16] Zhou S, Li Z, Li X, Ye Y, Wang M, Jiang J, et al. Crosstalk between endothelial cells and dermal papilla entails hair regeneration and angiogenesis during aging. J Adv Res 2024; 70: 339-53. https://doi.org/10.1016/j.jare.2024.05.006.

[17] Lei M, Schumacher LJ, Lai Y-C, Juan W-T, Yeh C-Y, Wu P, et al. Self-organization process in newborn skin organoid formation inspires strategy to restore hair regeneration of adult cells. Proc Natl Acad Sci 2017; 114: E7101-10. https://doi.org/10.1073/pnas.1700475114.

[18] Gao Q, Wang J, Zhang H, Wang J, Jing Y, Su J. Organoid vascularization: strategies and applications. Adv Healthc Mater 2025; 14: e2500301. https://doi.org/10.1002/adhm.202500301.

[19] Rouwkema J, Khademhosseini A. Vascularization and angiogenesis in tissue engineering: beyond creating static networks. Trends Biotechnol 2016; 34: 733-45. https://doi.org/10.1016/j.tibtech.2016.03.002.

[20] Li Z, Yu D, Zhou C, Wang F, Lu K, Liu Y, et al. Engineering vascularised organoid-on-a-chip: strategies, advances and future perspectives. Biomater Transl 2024; 5: 21-32. https://doi.org/10.12336/biomatertransl.2024.01.003.

[21] Jun Y, Nguyen-Ngoc K-V, Sai S, Bender RHF, Gong W, Kravets V, et al. Engineered vasculature induces functional maturation of pluripotent stem cell-derived islet organoids. Dev Cell 2025; 60: 2455-2469.e7. https://doi.org/10.1016/j.devcel.2025.04.024.

[22] Sun X-Y, Ju X-C, Li Y, Zeng P-M, Wu J, Zhou Y-Y, et al. Generation of vascularized brain organoids to study neurovascular interactions. Elife 2022; 11: e76707. https://doi.org/10.7554/eLife.76707.

[23] Fumadó Navarro J, Crilly S, Chan WK, Browne S, Mason JO, Vallejo-Giraldo C, et al. Cerebral organoids with integrated endothelial networks emulate the neurovascular unit and mitigate core necrosis. Adv Sci (Weinh) 2025; 12: e07256. https://doi.org/10.1002/advs.202507256.

[24] Saiki N, Nio Y, Yoneyama Y, Kawamura S, Iwasawa K, Kawakami E, et al. Self-organization of sinusoidal vessels in pluripotent stem cell-derived human liver bud organoids. Nat Biomed Eng 2025; 9: 1869-85. https://doi.org/10.1038/s41551-025-01416-6.

[25] Zhu L, Liu S, Yang Z, Yang L, Yang Y, Han P, et al. Construction of vascularized liver microtissues recapitulates angiocrine-mediated hepatocytes maturation and enhances therapeutic efficacy for acute liver failure. Bioact Mater 2025; 50: 525-39. https://doi.org/10.1016/j.bioactmat.2025.04.030.

[26] Klinger D, Naftaly JA, Red-Horse K. Vascular organoids get a speed boost for regenerative repair. Cell Stem Cell 2025; 32: 1185-7. https://doi.org/10.1016/j.stem.2025.07.002.

[27] Gong L, Zhang Y, Zhu Y, Lee U, Luo AC, Li X, et al. Rapid generation of functional vascular organoids via simultaneous transcription factor activation of endothelial and mural lineages. Cell Stem Cell 2025; 32: 1200-1217.e6. https://doi.org/10.1016/j.stem.2025.05.014.

[28] Patsch C, Challet-Meylan L, Thoma EC, Urich E, Heckel T, O’Sullivan JF, et al. Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells. Nat Cell Biol 2015; 17: 994-1003. https://doi.org/10.1038/ncb3205.

[29] Wimmer RA, Leopoldi A, Aichinger M, Wick N, Hantusch B, Novatchkova M, et al. Human blood vessel organoids as a model of diabetic vasculopathy. Nature 2019; 565: 505-10. https://doi.org/10.1038/s41586-018-0858-8.

[30] Wimmer RA, Leopoldi A, Aichinger M, Kerjaschki D, Penninger JM. Generation of blood vessel organoids from human pluripotent stem cells. Nat Protoc 2019; 14: 3082-100. https://doi.org/10.1038/s41596-019-0213-z.

[31] Nwokoye PN, Abilez OJ. Bioengineering methods for vascularizing organoids. Cell Rep Methods 2024; 4: 100779. https://doi.org/10.1016/j.crmeth.2024.100779.

[32] Morita R, Suzuki M, Kasahara H, Shimizu N, Shichita T, Sekiya T, et al. ETS transcription factor ETV2 directly converts human fibroblasts into functional endothelial cells. Proc Natl Acad Sci USA 2015; 112: 160-5. https://doi.org/10.1073/pnas.1413234112.

[33] Lee S, Park C, Han JW, Kim JY, Cho K, Kim EJ, et al. Direct reprogramming of human dermal fibroblasts into endothelial cells using ER71/ETV2. Circ Res 2017; 120: 848-61. https://doi.org/10.1161/CIRCRESAHA.116.309833.

[34] Cakir B, Xiang Y, Tanaka Y, Kural MH, Parent M, Kang Y-J, et al. Engineering of human brain organoids with a functional vascular-like system. Nat Methods 2019; 16: 1169-75. https://doi.org/10.1038/s41592-019-0586-5.

[35] Grath A, Dai G. SOX17/ETV2 improves the direct reprogramming of adult fibroblasts to endothelial cells. Cell Rep Methods 2024; 4: 100732. https://doi.org/10.1016/j.crmeth.2024.100732.

[36] Gong L, Melero-Martin JM . Protocol for generating human vascular organoids via orthogonal activation of ETV2 and NKX3.1. STAR Protoc 2025; 7: 104294. https://doi.org/10.1016/j.xpro.2025.104294.

[37] Dailamy A, Parekh U, Katrekar D, Kumar A, McDonald D, Moreno A, et al. Programmatic introduction of parenchymal cell types into blood vessel organoids. Stem Cell Reports 2021; 16: 2432-41. https://doi.org/10.1016/j.stemcr.2021.08.014.

[38] Kshirsagar A, Mnatsakanyan H, Kulkarni S, Guo J, Cheng K, Ofria LD, et al. Multi-region brain organoids integrating cerebral, mid-hindbrain, and endothelial systems. Adv Sci (Weinh) 2025; 12: e03768. https://doi.org/10.1002/advs.202503768.

[39] Zhang C, Shen Y, Huang M, Wang G, Miao Q, Shi H, et al. Dynamic hydrogel mechanics in organoid engineering: from matrix design to translational paradigms. Bioact Mater 2026; 55: 144-70. https://doi.org/10.1016/j.bioactmat.2025.09.021.

[40] Tang Z, Xia Q, Li J, Chen J, Wu X, Li J, et al. Microenvironment-feedback hydrogel for precise sequential repair of acute infectious wounds. Biomed Tech 2025; 12: 100120. https://doi.org/10.1016/j.bmt.2025.100120.

[41] Han J, Halwachs K, West T, Larsen B, Sacks MS, Rosales AM, et al. Matrix stiffness regulates mechanotransduction and vascular network formation of hiPSC-derived endothelial progenitors encapsulated in 3D hydrogels. bioRxiv 2025.04.11.648340. https://doi.org/10.1101/2025.04.11.648340.

[42] Sun D, Zhang K, Zheng F, Yang G, Yang M, Xu Y, et al. Matrix viscoelasticity controls differentiation of human blood vessel organoids into arterioles and promotes neovascularization in myocardial infarction. Adv Mater 2025; 37: e2410802. https://doi.org/10.1002/adma.202410802.

[43] Wolff L, Hendrix S. Rethinking Matrigel: the complex journey to matrix alternatives in organoid culture. Adv Sci (Weinh) 2025; 12: e08734. https://doi.org/10.1002/advs.202508734.

[44] Brembilla N-C, El-Harane S, Durual S, Krause K-H, Preynat-Seauve O. Adipose-derived stromal cells exposed to RGD motifs enter an angiogenic stage regulating endothelial cells. Int J Mol Sci 2025; 26: 867. https://doi.org/10.3390/ijms26030867.

[45] Homan KA, Gupta N, Kroll KT, Kolesky DB, Skylar-Scott M, Miyoshi T, et al. Flow-enhanced vascularization and maturation of kidney organoids in vitro. Nat Methods 2019; 16: 255-62. https://doi.org/10.1038/s41592-019-0325-y.

[46] Wang Y, Guo Y, Wang P, Liu J, Zhang X, Liu Q, et al. An engineered human placental organoid microphysiological system in a vascular niche to model viral infection. Commun Biol 2025; 8: 669. https://doi.org/10.1038/s42003-025-08057-0.

[47] Shin YJ, Safina D, Zheng Y, Levenberg S. Microvascularization in 3D human engineered tissue and organoids. Annu Rev Biomed Eng 2025; 27: 473-98. https://doi.org/10.1146/annurev-bioeng-103023-115236.

[48] Nikolova MT, He Z, Seimiya M, Jonsson G, Cao W, Okuda R, et al. Fate and state transitions during human blood vessel organoid development. Cell 2025; 188: 3329-3348.e31. https://doi.org/10.1016/j.cell.2025.03.037.

[49] Thai VL, Ramos-Rodriguez DH, Mesfin M, Leach JK. Hydrogel degradation promotes angiogenic and regenerative potential of cell spheroids for wound healing. Mater Today Bio 2023; 22: 100769. https://doi.org/10.1016/j.mtbio.2023.100769.

[50] Lee J, Rabbani CC, Gao H, Steinhart MR, Woodruff BM, Pflum ZE, et al. Hair-bearing human skin generated entirely from pluripotent stem cells. Nature 2020; 582: 399-404. https://doi.org/10.1038/s41586-020-2352-3.

[51] Choudhury S, Dhoke NR, Chawla S, Das A. Bioengineered MSCCxcr2 transdifferentiated keratinocyte-like cell-derived organoid potentiates skin regeneration through ERK1/2 and STAT3 signaling in diabetic wound. Cell Mol Life Sci 2024; 81: 172. https://doi.org/10.1007/s00018-023-05057-3.

[52] Chen Z, Bo Q, Wang C, Xu Y, Fei X, Chen R. Single BMSC-derived cartilage organoids for gradient heterogeneous osteochondral regeneration by leveraging native vascular microenvironment. J Nanobiotechnol 2025; 23: 325. https://doi.org/10.1186/s12951-025-03403-0.

[53] Ji Y, Chen X, Wang Z, Meek CJ, McLean JL, Yang Y, et al. Alzheimer’s disease patient brain extracts induce multiple pathologies in novel vascularized neuroimmune organoids for disease modeling and drug discovery. Mol Psychiatry 2025; 30: 4558-75. https://doi.org/10.1038/s41380-025-03041-w.

[54] Sun C, Wu G, Wu D, Wu L, Lu Q, Hu W, et al. Determining the optimal transplantation window in hepatic organoids via real-time biosensing of vascularization and metabolic maturation utilizing the integrated organoid-on-a-chip platform. Biosens Bioelectron 2026; 292: 118057. https://doi.org/10.1016/j.bios.2025.118057.

[55] Li S, Kan F, Ji S, Cao D, Liu H, Wu J. Development of vascularized adipose organoids from PBMSCs and their application in evaluating Celastrol’s effects. Adipocyte 2025; 14: 2548787. https://doi.org/10.1080/21623945.2025.2548787.

[56] Yi S, Huang M, Xian C, Kong X, Yin S, Peng J, et al. Single-cell transcriptomics of vascularized human brain organoids decipher lineage-specific stress adaptation in fetal hypoxia-reoxygenation injury. Theranostics 2025; 15: 7001-24. https://doi.org/10.7150/thno.117001.

[57] Jgamadze D, Lim JT, Zhang Z, Harary PM, Germi J, Mensah-Brown K, et al. Structural and functional integration of human forebrain organoids with the injured adult rat visual system. Cell Stem Cell 2023; 30: 137-152.e7. https://doi.org/10.1016/j.stem.2023.01.004.

[58] Ondriš J, Schwamborn JC, Olcese U. Functional recovery through the plastic adaptation of organoid grafts in cortical tissue. Cell Mol Life Sci 2025; 82: 227. https://doi.org/10.1007/s00018-025-05767-w.

[59] Song S, Zhang J, Fang Y, Li W, Zeng H, Fang Z, et al. Nerve-bone crosstalk manipulates bone organoid development and bone regeneration: a review and perspectives. Organoid Res 2025; 1: 8294. https://doi.org/10.36922/or.8294.

[60] Zhang H, Qin C, Zhang M, Han Y, Ma J, Wu J, et al. Calcium silicate nanowires-containing multicellular bioinks for 3D bioprinting of neural-bone constructs. Nano Today 2022; 46: 101584. https://doi.org/10.1016/j.nantod.2022.101584.

[61] Wang T, Li W, Zhang Y, Xu X, Qiang L, Miao W, et al. Bioprinted constructs that simulate nerve-bone crosstalk to improve microenvironment for bone repair. Bioact Mater 2023; 27: 377-93. https://doi.org/10.1016/j.bioactmat.2023.02.013.

[62] Zhang H, Zhang M, Zhai D, Qin C, Wang Y, Ma J, et al. Polyhedron-like biomaterials for innervated and vascularized bone regeneration. Adv Mater 2023; 35: e2302716. https://doi.org/10.1002/adma.202302716.

[63] Faustino Martins J-M, Fischer C, Urzi A, Vidal R, Kunz S, Ruffault P-L, et al. Self-organizing 3D human trunk neuromuscular organoids. Cell Stem Cell 2020; 26: 172-186.e6. https://doi.org/10.1016/j.stem.2019.12.007.

[64] Zhang M, Qin C, Wang Y, Hu X, Ma J, Zhuang H, et al. 3D printing of tree-like scaffolds for innervated bone regeneration. Addit Manuf 2022; 54: 102721. https://doi.org/10.1016/j.addma.2022.102721.

[65] Wang J, Yang Q, Saiding Q, Chen L, Liu M, Wang Z, et al. Geometric angles and gene expression in cells for structural bone regeneration. Adv Sci (Weinh) 2023; 10: e2304111. https://doi.org/10.1002/advs.202304111.

[66] Workman MJ, Mahe MM, Trisno S, Poling HM, Watson CL, Sundaram N, et al. Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system. Nat Med 2017; 23: 49-59. https://doi.org/10.1038/nm.4233.

[67] Afshar, Bakooshli M, Lippmann ES, Mulcahy B, Iyer N, Nguyen CT, Tung K, et al. A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction. Elife 2019; 8: e44530. https://doi.org/10.7554/eLife.44530.

[68] Guo S, He C. Bioprinted scaffold remodels the neuromodulatory microenvironment for enhancing bone regeneration. Adv Funct Mater 2023; 33: 2304172. https://doi.org/10.1002/adfm.202304172.

[69] Hou P, Chen S, Tuerxun M, Wang S, Yan W, Tang J. Neuralized and vascularized fast bone regeneration using recombinant humanized type 1 collagen and native bone composite inorganic salts. APL Mater 2025; 13: 031103. https://doi.org/10.1063/5.0257593.

[70] Andersen J, Revah O, Miura Y, Thom N, Amin ND, Kelley KW, et al. Generation of functional human 3D cortico-motor assembloids. Cell 2020; 183: 1913-1929.e26. https://doi.org/10.1016/j.cell.2020.11.017.

[71] Kitahara T, Sakaguchi H, Morizane A, Kikuchi T, Miyamoto S, Takahashi J. Axonal extensions along corticospinal tracts from transplanted human cerebral organoids. Stem Cell Rep 2020; 15: 467-81. https://doi.org/10.1016/j.stemcr.2020.06.016.

[72] Cao S-Y, Yang D, Huang Z-Q, Lin Y-H, Wu H-Y, Chang L, et al. Cerebral organoids transplantation repairs infarcted cortex and restores impaired function after stroke. NPJ Regen Med 2023; 8: 27. https://doi.org/10.1038/s41536-023-00301-7.

[73] Zheng X, Han D, Liu W, Wang X, Pan N, Wang Y, et al. Human iPSC-derived midbrain organoids functionally integrate into striatum circuits and restore motor function in a mouse model of Parkinson’s disease. Theranostics 2023; 13: 2673-92. https://doi.org/10.7150/thno.80271.

[74] Xu J, Fang S, Deng S, Li H, Lin X, Huang Y, et al. Generation of neural organoids for spinal-cord regeneration via the direct reprogramming of human astrocytes. Nat Biomed Eng 2022; 7: 253-69. https://doi.org/10.1038/s41551-022-00963-6.

[75] Fang Y-T, Lin Y-T, Tseng W-L, Tseng P, Hua G-L, Chao Y-J, et al. Neuroimmunomodulation of vagus nerve stimulation and the therapeutic implications. Front Aging Neurosci 2023; 15: 1173987. https://doi.org/10.3389/fnagi.2023.1173987.

[76] Nava RG, Adri AS, Filgueiras IS, Nóbile AL, Barcelos PM, Corrêa YLG, et al. Modulation of neuroimmune cytokine networks by antidepressants: implications in mood regulation. Transl Psychiatry 2025; 15: 314. https://doi.org/10.1038/s41398-025-03532-y.

[77] Bogoslowski A, An M, Penninger JM. Incorporating immune cells into organoid models: essential for studying human disease. Organoids 2023; 2: 140-55. https://doi.org/10.3390/organoids2030011.

[78] Neves JF. Expanding organoid complexity to advance immunology research. Clin Exp Immunol 2024; 218: 14-5. https://doi.org/10.1093/cei/uxae067.

[79] Schmidt-Bleek K, Kwee BJ, Mooney DJ, Duda GN . Boon and bane of inflammation in bone tissue regeneration and its link with angiogenesis. Tissue Eng Part B Rev 2015; 21: 354-64. https://doi.org/10.1089/ten.TEB.2014.0677.

[80] Li Z, Zhou S, Ye Y, Li K, Liang H, Wang N, et al. Low temperatures promote the healing of skin micro-injuries by inhibiting IL1β. Regenesis Repair Rehabil 2025; 1: 6-15. https://doi.org/10.1016/j.rerere.2024.09.001.

[81] Caballero-Sánchez N, Alonso-Alonso S, Nagy L. Regenerative inflammation: when immune cells help to re-build tissues. FEBS J 2024; 291: 1597-614. https://doi.org/10.1111/febs.16693.

[82] Ziemkiewicz N, Hilliard G, Pullen NA, Garg K. The role of innate and adaptive immune cells in skeletal muscle regeneration. Int J Mol Sci 2021; 22: 22. https://doi.org/10.3390/ijms22063265.

[83] Wilgus TA. Immune cells in the healing skin wound: influential players at each stage of repair. Pharmacol Res 2008; 58: 112-6. https://doi.org/10.1016/j.phrs.2008.07.009.

[84] Eming SA, Hammerschmidt M, Krieg T, Roers A. Interrelation of immunity and tissue repair or regeneration. Semin Cell Dev Biol 2009; 20: 517-27. https://doi.org/10.1016/j.semcdb.2009.04.009.

[85] Wilgus TA. Inflammation as an orchestrator of cutaneous scar formation: a review of the literature. Plast Aesthet Res 2020; 7: 54. https://doi.org/10.20517/2347-9264.2020.150.

[86] Lavine KJ, Epelman S, Uchida K, Weber KJ, Nichols CG, Schilling JD, et al. Distinct macrophage lineages contribute to disparate patterns of cardiac recovery and remodeling in the neonatal and adult heart. Proc Natl Acad Sci USA 2014; 111: 16029-34. https://doi.org/10.1073/pnas.1406508111.

[87] Burzyn D, Kuswanto W, Kolodin D, Shadrach JL, Cerletti M, Jang Y, et al. A special population of regulatory T cells potentiates muscle repair. Cell 2013; 155: 1282-95. https://doi.org/10.1016/j.cell.2013.10.054.

[88] Jiang S, Deng T, Cheng H, Liu W, Shi D, Yuan J, et al. Macrophage-organoid co-culture model for identifying treatment strategies against macrophage-related gemcitabine resistance. J Exp Clin Cancer Res 2023; 42: 199. https://doi.org/10.1186/s13046-023-02756-4.

[89] Li P, Pachis ST, Xu G, Schraauwen R, Incitti R, de Vries AC, et al. Mpox virus infection and drug treatment modelled in human skin organoids. Nat Microbiol 2023; 8: 2067-79. https://doi.org/10.1038/s41564-023-01489-6.

[90] Wagar LE, Salahudeen A, Constantz CM, Wendel BS, Lyons MM, Mallajosyula V, et al. Modeling human adaptive immune responses with tonsil organoids. Nat Med 2021; 27: 125-35. https://doi.org/10.1038/s41591-020-01145-0.

[91] van den Bogaard EH, Tjabringa GS, Joosten I, Vonk-Bergers M, van Rijssen E, Tijssen HJ, et al. Crosstalk between keratinocytes and T cells in a 3D microenvironment: a model to study inflammatory skin diseases. J Invest Dermatol 2014; 134: 719-27. https://doi.org/10.1038/jid.2013.417.

[92] Vazquez-Armendariz AI, Heiner M, El Agha E, Salwig I, Hoek A, Hessler MC, et al. Multilineage murine stem cells generate complex organoids to model distal lung development and disease. EMBO J 2020; 39: e103476. https://doi.org/10.15252/embj.2019103476.

[93] Liu X, Yang J, Yan Y, Li Q, Huang R-L. Unleashing the potential of adipose organoids: a revolutionary approach to combat obesity-related metabolic diseases. Theranostics 2024; 14: 2075-98. https://doi.org/10.7150/thno.93919.

[94] Kwak BS, Jin S-P, Kim SJ, Kim EJ, Chung JH, Sung JH. Microfluidic skin chip with vasculature for recapitulating the immune response of the skin tissue. Biotechnol Bioeng 2020; 117: 1853-63. https://doi.org/10.1002/bit.27320.

[95] Kim J, Guhl S, Babina M, Zuberbier T, Artuc M. Integration of the human dermal mast cell into the organotypic co-culture skin model. Methods Mol Biol 2020; 2163: 91-107. https://doi.org/10.1007/978-1-0716-0696-4_8.

[96] Jowett GM, Read E, Roberts LB, Coman D, Vilà González M, Zabinski T, et al. Organoids capture tissue-specific innate lymphoid cell development in mice and humans. Cell Rep 2022; 40: 111281. https://doi.org/10.1016/j.celrep.2022.111281.

[97] Frenz-Wiessner S, Fairley SD, Buser M, Goek I, Salewskij K, Jonsson G, et al. Generation of complex bone marrow organoids from human induced pluripotent stem cells. Nat Methods 2024; 21: 868-81. https://doi.org/10.1038/s41592-024-02172-2.

[98] Shankar AS, Tejeda-Mora H, Du Z, Nlandu Q, Palomares-Cabeza V, van den Bosch TPP, et al. Interactions of the immune system with human kidney organoids. Transpl Int 2024; 37: 12468. https://doi.org/10.3389/ti.2024.12468.

[99] Papadimitropoulos A, Scherberich A, Güven S, Theilgaard N, Crooijmans HJA, Santini F, et al. A 3D in vitro bone organ model using human progenitor cells. Eur Cell Mater 2011; 21: 445-58; discussion 458. https://doi.org/10.22203/ecm.v021a33.

[100] Kluger MS. Vascular endothelial cell adhesion and signaling during leukocyte recruitment. Adv Dermatol 2004; 20: 163-201.

[101] Mueller SN. Neural control of immune cell trafficking. J Exp Med 2022; 219: e20211604. https://doi.org/10.1084/jem.20211604.

[102] Udit S, Blake K, Chiu IM. Somatosensory and autonomic neuronal regulation of the immune response. Nat Rev Neurosci 2022; 23: 157-71. https://doi.org/10.1038/s41583-021-00555-4.

[103] Madl CM, Heilshorn SC, Blau HM. Bioengineering strategies to accelerate stem cell therapeutics. Nature 2018; 557: 335-42. https://doi.org/10.1038/s41586-018-0089-z.

[104] Mostina M, Sun J, Sim SL, Ahmed IA, Souza-Fonesca-Guimaraes F, Wolvetang EJ, et al. Coordinated development of immune cell populations in vascularized skin organoids from human induced pluripotent stem cells. Adv Healthc Mater 2025; 14: e02108. https://doi.org/10.1002/adhm.202502108.

[105] Ahn S-J, Lee S, Kwon D, Oh S, Park C, Jeon S, et al. Essential guidelines for manufacturing and application of organoids. Int J Stem Cells 2024; 17: 102-12. https://doi.org/10.15283/ijsc24047.

[106] van Zanten J, Jorritsma-Smit A, Westra H, Baanstra M, de Bruin-Jellema A, Allersma D, et al. Optimization of the production process of clinical-grade human salivary gland organoid-derived cell therapy for the treatment of radiation-induced xerostomia in head and neck cancer. Pharmaceutics 2024; 16: 435. https://doi.org/10.3390/pharmaceutics16030435.

[107] Ingber DE. Human organs-on-chips for disease modelling, drug development and personalized medicine. Nat Rev Genet 2022; 23: 467-91. https://doi.org/10.1038/s41576-022-00466-9.

[108] Schurink IJ, de Jonge J, van der Laan LJW. Organoid technology starts to deliver: repairing damaged liver grafts during normothermic machine perfusion. Transplantation 2021; 105: 1886-7. https://doi.org/10.1097/TP.0000000000003790.