维系类器官培养稳定性的相关研究进展
Research Progress on Maintaining the Stability of Organoid Culture
DOI: 10.12677/acm.2025.1592554, PDF,   
作者: 夏雪妍:西安医学院研究生工作部,陕西 西安;段宝军*:陕西省人民医院肿瘤内科,陕西 西安
关键词: 类器官培养稳定性血管化Organoid Culture Stability Vascularization
摘要: 类器官作为三维微型器官模型,能够高度模拟体内器官的结构与功能,在疾病建模、药物筛选等领域彰显出巨大潜力。不过,其培养过程中存在的稳定性难题,极大地限制了实际应用。类器官的稳定性体现在两个维度:一方面是培养环境的稳态保持,涉及基质构成、生长因子、机械刺激等外源性要素;另一方面是细胞的内在稳态,包含遗传稳定性、代谢均衡以及细胞异质性等内源性要素。本文全面综述了维持类器官培养环境及内源稳定性的前沿手段,包括工程化基质、血管化技术、标准化流程、菌群平衡等方面,通过批判性评估各技术的优劣提出整合性策略,并对其临床转化前景进行了展望。
Abstract: As three-dimensional miniaturized organ models, organoids faithfully replicate the structural and functional characteristics of native tissues, demonstrating substantial potential in disease modeling, drug screening, and related fields. However, cultivation stability challenges significantly hinder their practical applications. Organoid stability encompasses two dimensions: Extrinsic homeostasis, maintenance of the culture environment through exogenous factors (e.g. matrix composition, growth factors, mechanical stimuli); Intrinsic cellular homeostasis, preservation of endogenous properties including genetic stability, metabolic equilibrium, and cellular heterogeneity. This review comprehensively examines cutting-edge strategies for sustaining both environmental and cellular stability in organoid cultures, covering engineered matrices, vascularization techniques, standardized protocols, and microbial community regulation. Future prospects for clinical translation are also discussed.
文章引用:夏雪妍, 段宝军. 维系类器官培养稳定性的相关研究进展[J]. 临床医学进展, 2025, 15(9): 771-777. https://doi.org/10.12677/acm.2025.1592554

参考文献

[1] Marsee, A., Roos, F.J.M., Verstegen, M.M.A., Gehart, H., de Koning, E., Lemaigre, F., et al. (2021) Building Consensus on Definition and Nomenclature of Hepatic, Pancreatic, and Biliary Organoids. Cell Stem Cell, 28, 816-832. [Google Scholar] [CrossRef] [PubMed]
[2] Yan, H.H.N., Chan, A.S., Lai, F.P. and Leung, S.Y. (2023) Organoid Cultures for Cancer Modeling. Cell Stem Cell, 30, 917-937. [Google Scholar] [CrossRef] [PubMed]
[3] Sauter, M.M., Noel, H.R., Sinha, D., Nelson, E.C., Xiong, M.N., Gamm, D.M., et al. (2025) AAV2.7m8 Transduction of Stage 2 Human Retinal Organoids Induces Highly Variable Responses in Innate and Inflammatory Gene Expression and Cytokine Secretion. Experimental Eye Research, 258, Article ID: 110478. [Google Scholar] [CrossRef] [PubMed]
[4] Baghdadi, M.B., Houtekamer, R.M., Perrin, L., Rao-Bhatia, A., Whelen, M., Decker, L., et al. (2024) Piezo-Dependent Mechanosensing Is Essential for Intestinal Stem Cell Fate Decision and Maintenance. Science, 386, eadj7615. [Google Scholar] [CrossRef] [PubMed]
[5] Wijnakker, J.J.A.P.M., van Son, G.J.F., Krueger, D., van de Wetering, W.J., Lopez-Iglesias, C., Schreurs, R., et al. (2024) Integrin-Activating Yersinia Protein Invasin Sustains Long-Term Expansion of Primary Epithelial Cells as 2D Organoid Sheets. Proceedings of the National Academy of Sciences of the United States of America, 122, e2420595121. [Google Scholar] [CrossRef] [PubMed]
[6] Chalard, A.E., Dixon, A.W., Taberner, A.J. and Malmström, J. (2022) Visible-light Stiffness Patterning of Gelma Hydrogels Towards in Vitro Scar Tissue Models. Frontiers in Cell and Developmental Biology, 10, Article 946754. [Google Scholar] [CrossRef] [PubMed]
[7] Rijal, G. and Li, W. (2017) A Versatile 3D Tissue Matrix Scaffold System for Tumor Modeling and Drug Screening. Science Advances, 3, e1700764. [Google Scholar] [CrossRef] [PubMed]
[8] Saiki, N., Nio, Y., Yoneyama, Y., Kawamura, S., Iwasawa, K., Kawakami, E., Araki, K., Fukumura, J., Sakairi, T., Kono, T., et al. (2024) Self-Organization of Sinusoidal Vessels in Pluripotent Stem Cell-Derived Human Liver Bud Organoids. Cold Spring Harbor Laboratory. [Google Scholar] [CrossRef] [PubMed]
[9] Cakir, B., Xiang, Y., Tanaka, Y., Kural, M.H., Parent, M., Kang, Y., et al. (2019) Engineering of Human Brain Organoids with a Functional Vascular-Like System. Nature Methods, 16, 1169-1175. [Google Scholar] [CrossRef] [PubMed]
[10] Zhang, X., Jiang, W., Wu, X., Xie, C., Zhang, Y., Li, L., et al. (2025) Divide-and-Conquer Strategy with Engineered Ossification Center Organoids for Rapid Bone Healing through Developmental Cell Recruitment. Nature Communications, 16, Article No. 6200. [Google Scholar] [CrossRef] [PubMed]
[11] Ingber, D.E. (1997) Tensegrity: The Architectural Basis of Cellular Mechanotransduction. Annual Review of Physiology, 59, 575-599. [Google Scholar] [CrossRef] [PubMed]
[12] Meng, F., Shen, C., Yang, L., Ni, C., Huang, J., Lin, K., et al. (2022) Mechanical Stretching Boosts Expansion and Regeneration of Intestinal Organoids through Fueling Stem Cell Self-Renewal. Cell Regeneration, 11, Article No. 39. [Google Scholar] [CrossRef] [PubMed]
[13] Zhou, L., Shi, Z., Yang, X., Zeng, J., You, Z., Zhang, Y., et al. (2025) Tension-Induced Directional Migration of Hepatic Stellate Cells Potentially Coordinates Liver Fibrosis Progression. Nature Biomedical Engineering. [Google Scholar] [CrossRef] [PubMed]
[14] Usman, O.H., Zhang, L., Xie, G., Kocher, H.M., Hwang, C., Wang, Y.J., et al. (2022) Genomic Heterogeneity in Pancreatic Cancer Organoids and Its Stability with Culture. NPJ Genomic Medicine, 7, Article No. 71. [Google Scholar] [CrossRef] [PubMed]
[15] Klaasen, S.J., Truong, M.A., van Jaarsveld, R.H., Koprivec, I., Štimac, V., de Vries, S.G., et al. (2022) Nuclear Chromosome Locations Dictate Segregation Error Frequencies. Nature, 607, 604-609. [Google Scholar] [CrossRef] [PubMed]
[16] Dekkers, J.F., van Vliet, E.J., Sachs, N., Rosenbluth, J.M., Kopper, O., Rebel, H.G., et al. (2021) Long-Term Culture, Genetic Manipulation and Xenotransplantation of Human Normal and Breast Cancer Organoids. Nature Protocols, 16, 1936-1965. [Google Scholar] [CrossRef] [PubMed]
[17] Koch, L.S., Choy Buentello, D. and Broersen, K. (2023) Robust Tissue Fabrication for Long-Term Culture of iPSC-Derived Brain Organoids for Aging Research. Journal of Visualized Experiments, 195, e64586. [Google Scholar] [CrossRef] [PubMed]
[18] Cai, H., Tian, C., Chen, L., Yang, Y., Sun, A.X., McCracken, K., et al. (2025) Vascular Network-Inspired Diffusible Scaffolds for Engineering Functional Midbrain Organoids. Cell Stem Cell, 32, 824-837.e5. [Google Scholar] [CrossRef] [PubMed]
[19] Hou, Q., Jia, J., Lin, J., Zhu, L., Xie, S., Yu, Q., et al. (2022) Bacillus Subtilis Programs the Differentiation of Intestinal Secretory Lineages to Inhibit Salmonella Infection. Cell Reports, 40, Article ID: 111416. [Google Scholar] [CrossRef] [PubMed]
[20] Zhang, J., Hernandez-Gordillo, V., Trapecar, M., Wright, C., Taketani, M., Schneider, K., et al. (2021) Coculture of Primary Human Colon Monolayer with Human Gut Bacteria. Nature Protocols, 16, 3874-3900. [Google Scholar] [CrossRef] [PubMed]
[21] Yao, W., Song, W., Deng, X., Lin, Y., Meng, R., Wang, J., et al. (2024) Harnessing the Engineered Probiotic‐Nanosystem to Remodulate Tumor Extracellular Matrix and Regulate Tumor‐Colonizing Bacteria for Improving Pancreatic Cancer Chemo‐Immunotherapy. Small, 21, e2406837. [Google Scholar] [CrossRef] [PubMed]
[22] Volta, V., Pérez-Baos, S., de la Parra, C., Katsara, O., Ernlund, A., Dornbaum, S., et al. (2021) A DAP5/eIF3d Alternate mRNA Translation Mechanism Promotes Differentiation and Immune Suppression by Human Regulatory T Cells. Nature Communications, 12, Article No. 6979. [Google Scholar] [CrossRef] [PubMed]
[23] Smith, T.J., Sundarraman, D., Melancon, E., Desban, L., Parthasarathy, R. and Guillemin, K. (2023) A Mucin-Regulated Adhesin Determines the Spatial Organization and Inflammatory Character of a Bacterial Symbiont in the Vertebrate Gut. Cell Host & Microbe, 31, 1371-1385.e6. [Google Scholar] [CrossRef] [PubMed]
[24] Tan, J.K., Macia, L. and Mackay, C.R. (2023) Dietary Fiber and SCFAs in the Regulation of Mucosal Immunity. Journal of Allergy and Clinical Immunology, 151, 361-370. [Google Scholar] [CrossRef] [PubMed]
[25] Yao, N., Jing, N., Lin, J., Niu, W., Yan, W., Yuan, H., et al. (2025) Patient-Derived Tumor Organoids for Cancer Immunotherapy: Culture Techniques and Clinical Application. Investigational New Drugs, 43, 394-404. [Google Scholar] [CrossRef] [PubMed]
[26] Yang, R., Qi, Y., Zhang, X., Gao, H. and Yu, Y. (2024) Living Biobank: Standardization of Organoid Construction and Challenges. Chinese Medical Journal, 137, 3050-3060. [Google Scholar] [CrossRef] [PubMed]
[27] Wang, X., Xia, T., Tang, H., Liu, X., Han, R., Zou, X., et al. (2022) Establishment of a Patient-Derived Organoid Model and Living Biobank for Nasopharyngeal Carcinoma. Annals of Translational Medicine, 10, 526-526. [Google Scholar] [CrossRef] [PubMed]
[28] Gong, S., He, K., Liu, Y., Luo, X., Ashraf, K., He, J., et al. (2025) Scalable Matrigel‐Free Suspension Culture for Generating High‐Quality Human Liver Ductal Organoids. Cell Proliferation, 58, e70033. [Google Scholar] [CrossRef] [PubMed]
[29] Ong, H.T., Karatas, E., Poquillon, T., Grenci, G., Furlan, A., Dilasser, F., et al. (2025) Digitalized Organoids: Integrated Pipeline for High-Speed 3D Analysis of Organoid Structures Using Multilevel Segmentation and Cellular Topology. Nature Methods, 22, 1343-1354. [Google Scholar] [CrossRef] [PubMed]
[30] Hong, F., Wang, X., Zhong, N., Zhang, Z., Lin, S., Zhang, M., et al. (2025) The Critical Role of BMP Signaling in Gastric Epithelial Cell Differentiation Revealed by Organoids. Cell Regeneration, 14, Article No. 18. [Google Scholar] [CrossRef] [PubMed]
[31] Hofer, M., Kim, Y., Broguiere, N., Gorostidi, F., Klein, J.A., Amieva, M.R., et al. (2025) Accessible Homeostatic Gastric Organoids Reveal Secondary Cell Type-Specific Host-Pathogen Interactions in Helicobacter pylori Infections. Nature Communications, 16, Article No. 2767. [Google Scholar] [CrossRef] [PubMed]
[32] 中华医学会消化病学分会医工交叉协作组. 中国经内镜消化系统常见恶性肿瘤组织取样及类器官培养专家共识(2024, 成都) [J]. 中华消化内镜杂志, 2024, 41(5): 337-350.
[33] Liu, J., Wu, G., Wu, D., Wu, L., Sun, C., Zhang, W., et al. (2025) Microfluidic Organoid-Slice-On-A-Chip System for Studying Anti-Cholangiocarcinoma Drug Efficacy and Hepatorenal Toxicity. Lab on a Chip, 25, 2839-2850. [Google Scholar] [CrossRef] [PubMed]
[34] Huang, Y., Liu, T., Huang, Q. and Wang, Y. (2024) From Organ-On-A-Chip to Human-On-A-Chip: A Review of Research Progress and Latest Applications. ACS Sensors, 9, 3466-3488. [Google Scholar] [CrossRef] [PubMed]
[35] He, C., Kalafut, N.C., Sandoval, S.O., Risgaard, R., Sirois, C.L., Yang, C., et al. (2023) BOMA, a Machine-Learning Framework for Comparative Gene Expression Analysis across Brains and Organoids. Cell Reports Methods, 3, Article ID: 100409. [Google Scholar] [CrossRef] [PubMed]
[36] Zheng, C., Wang, P., Zhang, D., Fang, Z., Feng, Y., Chen, J., et al. (2025) A Novel Organoid Model Retaining the Glioma Microenvironment for Personalized Drug Screening and Therapeutic Evaluation. Bioactive Materials, 53, 205-217. [Google Scholar] [CrossRef] [PubMed]