Stem Cell-Derived Organoids of the Pancreas: Evaluation of Endocrine and Exocrine Modeling Platforms
Publication: Tissue Engineering Part B: Reviews
Abstract
As miniature, three-dimensional emulates of individual human organs generated in vitro, organoids are increasingly recognized as complex, humanized models of development, disease, diagnostics, and drug discovery. Organoids exhibit organ-specific architecture, function, and multicellular composition, can be infinitely derived from pluripotent stem cells, and can be further directed toward organoids of the endocrine or exocrine pancreas. Pancreatic endocrine organoids are rapidly redefining diabetes therapies due to their ability to recapitulate glucose-responsive insulin secretion. Conversely, there is less focus on pancreatic exocrine organoids, which possess untapped potential for investigating disorders such as cancer and cystic fibrosis. This review first summarizes human pancreatic organogenesis to contextualize relevant differentiation pathways, then details protocols that guide human pluripotent stem cells through key developmental stages. Methods to enhance cellular maturation and establish higher-performing end products, as well as the therapeutic value of different pancreatic genres, are assessed. Furthermore, crucial gaps are identified, including limited insight into non-beta-endocrine cells, progenitor lineage bias, and off-target differentiation. By chronicling the advancements of all pancreatic organoid classes, the importance of creating more intricate constructs is underscored, which could lead to their broader application.
Abstract
Impact Statement
This review consolidates foundational work and recent advances in the generation and engineering of pancreatic organoids from human pluripotent stem cells, providing a comprehensive analysis of both endocrine and exocrine cell lineage development. Current differentiation protocols and the associated engineering innovations that improve the maturity of stem cell derivatives are evaluated. The review underscores the growing potential of organoids in disease modeling, drug discovery, and regenerative medicine. Emphasizing the need to expand the functional scope of organoids beyond β-cell-centric models is crucial for future efforts to develop physiologically complex and more clinically relevant pancreatic tissue equivalents.
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References
1. Scuderi S, Altobelli GG, Cimini V, et al. Cell-to-Cell adhesion and neurogenesis in human cortical development: A study comparing 2D monolayers with 3D organoid cultures. Stem Cell Reports 2021;16(2):264–280;
2. Cherubini A, Rusconi F, Piras R, et al. Exploring human pancreatic organoid modelling through single-cell RNA sequencing analysis. Commun Biol 2024;7(1):1527;
3. Mulaudzi PE, Abrahamse H, Crous A. Insights on three dimensional organoid studies for stem cell therapy in regenerative medicine. Stem Cell Rev Rep 2024;20(2):509–523;
4. Diane A, Mohammed LI, Al-Siddiqi HH. Islets in the body are never flat: Transitioning from two-dimensional (2D) monolayer culture to three-dimensional (3D) spheroid for better efficiency in the generation of functional hPSC-derived pancreatic β cells in vitro. Cell Commun Signal 2023;21(1):151;
5. Liu H, Wang Y, Cui K, et al. Advances in hydrogels in organoids and organs‐on‐a‐chip. Adv Mater 2019;31(50):e1902042;
6. Marsee A, Roos FJM, Verstegen MMA, et al.; HPB Organoid Consortium. Building consensus on definition and nomenclature of hepatic, pancreatic, and biliary organoids. Cell Stem Cell 2021;28(5):816–832;
7. Pagliuca FW, Millman JR, Gürtler M, et al. Generation of functional human pancreatic β cells in vitro. Cell 2014;159(2):428–439;
8. Breunig M, Merkle J, Wagner M, et al. Modeling plasticity and dysplasia of pancreatic ductal organoids derived from human pluripotent stem cells. Cell Stem Cell 2021;28(6):1105–1124.e19;
9. Frappart PO, Walter K, Gout J, et al. Pancreatic cancer-derived organoids—a disease modeling tool to predict drug response. United European Gastroenterol J 2020;8(5):594–606;
10. Hohwieler M, Illing A, Hermann PC, et al. Human pluripotent stem cell-derived acinar/ductal organoids generate human pancreas upon orthotopic transplantation and allow disease modelling. Gut 2017;66(3):473–486;
11. Dekkers JF, Wiegerinck CL, De Jonge HR, et al. A functional CFTR assay using primary cystic fibrosis intestinal organoids. Nat Med 2013;19(7):939–945;
12. Lee J, Snyder ER, Liu Y, et al. Reconstituting development of pancreatic intraepithelial neoplasia from primary human pancreas duct cells. Nat Commun 2017;8:14686;
13. Shih HP, Wang A, Sander M. Pancreas organogenesis: From lineage determination to morphogenesis. Annu Rev Cell Dev Biol 2013;29:81–105;
14. Mehta V, Hopson PE, Smadi Y, et al. Development of the human pancreas and its exocrine function. Front Pediatr 2022;10:909648;
15. Petersen MBK, Goncalves CAC, Kim YH, et al. Recapitulating and deciphering human pancreas development from human pluripotent stem cells in a dish. Curr Top Dev Biol 2018;129:143–190;
16. Leung PS. Overview of the Pancreas. Springer Netherlands; 2010. pp. 3–12.
17. Liu Y, Li N, Zhu Y. Pancreatic organoids: A frontier method for investigating pancreatic-related diseases. Int J Mol Sci 2023;24(4):4027;
18. Jiang L, Shen Y, Liu Y, et al. Making human pancreatic islet organoids: Progresses on the cell origins, biomaterials and three-dimensional technologies. Theranostics 2022;12(4):1537–1556;
19. Zhang X, Ma Z, Song E, et al. Islet organoid as a promising model for diabetes. Protein Cell 2022;13(4):239–257;
20. Cabrera O, Berman DM, Kenyon NS, et al. The unique cytoarchitecture of human pancreatic islets has implications for islet cell function. Proc Natl Acad Sci U S A 2006;103(7):2334–2339;
21. Moede T, Leibiger IB, Berggren P-O. Alpha cell regulation of beta cell function. Diabetologia 2020;63(10):2064–2075;
22. Arrojo E Drigo R, Jacob S, García-Prieto CF, et al. Structural basis for delta cell paracrine regulation in pancreatic islets. Nat Commun 2019;10(1):3700;
23. Hart NJ, Powers AC. Use of human islets to understand islet biology and diabetes: Progress, challenges and suggestions. Diabetologia 2019;62(2):212–222;
24. Pancreatic Polypeptide. The Pancreapedia: Exocrine Pancreas Knowledge Base;
25. Yoshihara E, O’Connor C, Gasser E, et al. Immune-evasive human islet-like organoids ameliorate diabetes. Nature 2020;586(7830):606–611;
26. Gamble A, Pepper AR, Bruni A, et al. The journey of islet cell transplantation and future development. Islets 2018;10(2):80–94;
27. Nair GG, Liu JS, Russ HA, et al. Recapitulating endocrine cell clustering in culture promotes maturation of human stem-cell-derived β cells. Nat Cell Biol 2019;21(2):263–274;
28. Balboa D, Barsby T, Lithovius V, et al. Functional, metabolic and transcriptional maturation of human pancreatic islets derived from stem cells. Nat Biotechnol 2022;40(7):1042–1055;
29. Jennings RE, Berry AA, Kirkwood-Wilson R, et al. Development of the human pancreas from foregut to endocrine commitment. Diabetes 2013;62(10):3514–3522;
30. Huang H, Bader TN, Jin S. Signaling molecules regulating pancreatic endocrine development from pluripotent stem cell differentiation. Int J Mol Sci 2020;21(16):5867;
31. D’Amour KA, Agulnick AD, Eliazer S, et al. Efficient differentiation of human embryonic stem cells to definitive endoderm. Nat Biotechnol 2005;23(12):1534–1541;
32. D’Amour KA, Bang AG, Eliazer S, et al. Production of pancreatic hormone–expressing endocrine cells from human embryonic stem cells. Nat Biotechnol 2006;24(11):1392–1401;
33. Russ HA, Parent AV, Ringler JJ, et al. Controlled induction of human pancreatic progenitors produces functional beta‐like cells in vitro. Embo J 2015;34(13):1759–1772;
34. Rezania A, Bruin JE, Arora P, et al. Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells. Nat Biotechnol 2014;32(11):1121–1133;
35. Velazco-Cruz L, Song J, Maxwell KG, et al. Acquisition of dynamic function in human stem cell-derived beta cells. Stem Cell Reports 2019;12(2):351–365;
36. McCracken KW, Wells JM. Molecular pathways controlling pancreas induction. Semin Cell Dev Biol 2012;23(6):656–662;
37. Gouon-Evans V, Boussemart L, Gadue P, et al. BMP-4 is required for hepatic specification of mouse embryonic stem cell–derived definitive endoderm. Nat Biotechnol 2006;24(11):1402–1411;
38. Green MD, Chen A, Nostro M-C, et al. Generation of anterior foregut endoderm from human embryonic and induced pluripotent stem cells. Nat Biotechnol 2011;29(3):267–272;
39. Bruin JE, Erener S, Vela J, et al. Characterization of polyhormonal insulin-producing cells derived in vitro from human embryonic stem cells. Stem Cell Res 2014;12(1):194–208;
40. Schaffer AE, Freude KK, Nelson SB, et al. Nkx6 transcription factors and Ptf1a function as antagonistic lineage determinants in multipotent pancreatic progenitors. Dev Cell 2010;18(6):1022–1029;
41. Gradwohl G, Dierich A, Lemeur M, et al. neurogenin3 is required for the development of the four endocrine cell lineages of the pancreas. Proc Natl Acad Sci U S A 2000;97(4):1607–1611;
42. Veres A, Faust AL, Bushnell HL, et al. Charting cellular identity during human in vitro β-cell differentiation. Nature 2019;569(7756):368–373;
43. Rezania A, Bruin JE, Riedel MJ, et al. Maturation of human embryonic stem cell-derived pancreatic progenitors into functional islets capable of treating pre-existing diabetes in mice. Diabetes 2012;61(8):2016–2029;
44. Chen S, Borowiak M, Fox JL, et al. A small molecule that directs differentiation of human ESCs into the pancreatic lineage. Nat Chem Biol 2009;5(4):258–265;
45. Konagaya S, Iwata H. Chemically defined conditions for long-term maintenance of pancreatic progenitors derived from human induced pluripotent stem cells. Sci Rep 2019;9(1):640;
46. Schulz TC, Young HY, Agulnick AD, et al. A scalable system for production of functional pancreatic progenitors from human embryonic stem cells. PLoS One 2012;7(5):e37004;
47. Millman JR, Xie C, Van Dervort A, et al. Generation of stem cell-derived β-cells from patients with type 1 diabetes. Nat Commun 2016;7(1):11463;
48. Roscioni SS, Migliorini A, Gegg M, et al. Impact of islet architecture on beta-cell heterogeneity, plasticity and function. Nat Rev Endocrinol 2016;12(12):695–709;
49. Alvarez-Dominguez JR, Melton DA. Cell maturation: Hallmarks, triggers, and manipulation. Cell 2022;185(2):235–249;
50. Barsby T, Ibrahim H, Lithovius V, et al. Differentiating functional human islet-like aggregates from pluripotent stem cells. STAR Protoc 2022;3(4):101711;
51. Wang W, Jin S, Ye K. Development of islet organoids from H9 human embryonic stem cells in biomimetic 3D scaffolds. Stem Cells Dev 2017;26(6):394–404;
52. Mamidi A, Prawiro C, Seymour PA, et al. Mechanosignalling via integrins directs fate decisions of pancreatic progenitors. Nature 2018;564(7734):114–118;
53. Greggio C, De Franceschi F, Figueiredo-Larsen M, et al. Artificial three-dimensional niches deconstruct pancreas developmentin vitro. Development 2013;140(21):4452–4462;
54. Candiello J, Grandhi TSP, Goh SK, et al. 3D heterogeneous islet organoid generation from human embryonic stem cells using a novel engineered hydrogel platform. Biomaterials 2018;177:27–39;
55. Richardson T, Kumta PN, Banerjee I. Alginate encapsulation of human embryonic stem cells to enhance directed differentiation to pancreatic islet-like cells. Tissue Eng Part A 2014;20(23–24):3198–3211;
56. Legøy TA, Vethe H, Abadpour S, et al. Encapsulation boosts islet-cell signature in differentiating human induced pluripotent stem cells via integrin signalling. Sci Rep 2020;10(1):414;
57. Hu M, Liu T, Huang H, et al. Extracellular matrix proteins refine microenvironments for pancreatic organogenesis from induced pluripotent stem cell differentiation. Theranostics 2025;15(6):2229–2249;
58. Yap WT, Salvay DM, Silliman MA, et al. Collagen IV-Modified scaffolds improve islet survival and function and reduce time to Euglycemia. Tissue Eng Part A 2013;19(21–22):2361–2372;
59. Bi H, Karanth SS, Ye K, et al. Decellularized tissue matrix enhances self-assembly of islet organoids from pluripotent stem cell differentiation. ACS Biomater Sci Eng 2020;6(7):4155–4165;
60. Patel SN, Ishahak M, Chaimov D, et al. Organoid microphysiological system preserves pancreatic islet function within 3D matrix. Sci Adv 2021;7(7):eaba5515;
61. Kim M, Cho S, Hwang DG, et al. Bioprinting of bespoke islet-specific niches to promote maturation of stem cell-derived islets. Nat Commun 2025;16(1):2205;
62. Huh D, Kim HJ, Fraser JP, et al. Microfabrication of human organs-on-chips. Nat Protoc 2013;8(11):2135–2157;
63. Bakhti M, Bottcher A, Lickert H. Modelling the endocrine pancreas in health and disease. Nat Rev Endocrinol 2019;15(3):155–171;
64. Tao T, Wang Y, Chen W, et al. Engineering human islet organoids from iPSCs using an organ-on-chip platform. Lab Chip 2019;19(6):948–958;
65. Mohammed JS, Wang Y, Harvat TA, et al. Microfluidic device for multimodal characterization of pancreatic islets. Lab Chip 2009;9(1):97–106;
66. Kopp JL, von Figura G, Mayes E, et al. Identification of Sox9-dependent acinar-to-ductal reprogramming as the principal mechanism for initiation of pancreatic ductal adenocarcinoma. Cancer Cell 2012;22(6):737–750;
67. Houbracken I, de Waele E, Lardon J, et al. Lineage tracing evidence for transdifferentiation of acinar to duct cells and plasticity of human pancreas. Gastroenterology 2011;141(2):731–741, 741 e1-4;
68. Baldan J, Houbracken I, Rooman I, et al. Adult human pancreatic acinar cells dedifferentiate into an embryonic progenitor-like state in 3D suspension culture. Sci Rep 2019;9(1):4040;
69. Mima A, Kimura A, Ito R, et al. Mechanistic elucidation of human pancreatic acinar development using single-cell transcriptome analysis on a human iPSC differentiation model. Sci Rep 2025;15(1):4668;
70. Huang L, Holtzinger A, Jagan I, et al. Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell– and patient-derived tumor organoids. Nat Med 2015;21(11):1364–1371;
71. Shirasawa S, Yoshie S, Yue F, et al. Pancreatic exocrine enzyme-producing cell differentiation via embryoid bodies from human embryonic stem cells. Biochem Biophys Res Commun 2011;410(3):608–613;
72. Aigha II, Abdelalim EM. NKX6.1 transcription factor: A crucial regulator of pancreatic beta cell development, identity, and proliferation. Stem Cell Res Ther 2020;11(1):459;
73. Turnpenny PD, Ellard S. Alagille syndrome: Pathogenesis, diagnosis and management. Eur J Hum Genet 2012;20(3):251–257;
74. Boocock GRB, Morrison JA, Popovic M, et al. Mutations in SBDS are associated with Shwachman–Diamond syndrome. Nat Genet 2003;33(1):97–101;
75. Freeman AJ, Ooi CY. Pancreatitis and pancreatic cystosis in cystic fibrosis. J Cyst Fibros 2017;16(Suppl 2):S79–S86;
76. Capurso G, Traini M, Piciucchi M, et al. Exocrine pancreatic insufficiency: Prevalence, diagnosis, and management. Clin Exp Gastroenterol 2019;12:129–139;
77. Mueller S, Engleitner T, Maresch R, et al. Evolutionary routes and KRAS dosage define pancreatic cancer phenotypes. Nature 2018;554(7690):62–68;
78. Boj SF, Hwang CI, Baker LA, et al. Organoid models of human and mouse ductal pancreatic cancer. Cell 2015;160(1–2):324–338;
79. Ferreira RMM, Sancho R, Messal HA, et al. Duct- and Acinar-Derived pancreatic ductal adenocarcinomas show distinct tumor progression and marker expression. Cell Rep 2017;21(4):966–978;
80. Tulpule A, Kelley JM, Lensch MW, et al. Pluripotent stem cell models of Shwachman-Diamond syndrome reveal a common mechanism for pancreatic and hematopoietic dysfunction. Cell Stem Cell 2013;12(6):727–736;
81. Gilbert MA, Spinner NB. Alagille syndrome: Genetics and Functional Models. Curr Pathobiol Rep 2017;5(3):233–241;
82. Ogawa M, Ogawa S, Bear CE, et al. Directed differentiation of cholangiocytes from human pluripotent stem cells. Nat Biotechnol 2015;33(8):853–861;
83. Sun H, Saeedi P, Karuranga S, et al. IDF diabetes atlas: Global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045. Diabetes Res Clin Pract 2022;183:109119;
84. Harris AR, Walker MJ, Gilbert F. Ethical and regulatory issues of stem cell-derived 3-dimensional organoid and tissue therapy for personalised regenerative medicine. BMC Med 2022;20(1):499;
85. Ameri J, Borup R, Prawiro C, et al. Efficient generation of glucose-responsive beta cells from isolated GP2(+) human pancreatic progenitors. Cell Rep 2017;19(1):36–49;
86. Merz S, Breunig M, Melzer MK, et al. Single-cell profiling of GP2-enriched pancreatic progenitors to simultaneously create acinar, ductal, and endocrine organoids. Theranostics 2023;13(6):1949–1973;
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