Research progress and future perspectives of tumor organoid

doi:10.19401/j.cnki.1007-3639.2024.08.006

Abstract

Abstract:

Tumor organoids, derived from tumor tissues, are three-dimensional structures created in vitro to mimic in vivo tumor growth. They possess comparable genetic lineage and pathological characteristics to the original tumor, making them a promising research platform for studying tumor biology. In recent years, significant advancements in techniques such as microfluidics, cell co-culture and vascularization have greatly promoted the development of Organ-on-a-Chip (OoC) and expanded the applications of tumor organoid models in both basic and clinical translational research. These advancements include leveraging tumor organoid models to investigate drug resistance mechanisms, perform drug screening, and facilitate clinical trials of precisional therapy. However, current organoid models still face several limitations, including the instability of cell culture quality, low cost-effectiveness, and the inability to accurately replicate the complex tumor microenvironment and spatial structures. Further research is needed to overcome these technical bottlenecks and improve the application of organoid models in cancer research, thereby further enhancing the depth of tumor research. This review summarized the development history and latest progress of tumor organoids. In terms of the latest applications of tumor organoids, this article reviewed disease modeling, research and development of innovative tumor drugs, and their application in personalized treatment. It also summarized the recent clinical researches of organoids; In addition, this review provided a detailed introduction to the technological progress of tumor organoids, including the development of novel culture devices, simulation of tumor microenvironment, and induction of angiogenesis. In summary, this review explored the latest developments, defects and future directions in the research of tumor organoids, aiming to provide reference for the study of tumor organoids.

Key words: Organoid, Organ-on-a-chip, Malignant tumor

CLC Number:

R73-3

XIAO Yi, WU Ming, YAO Gang. Research progress and future perspectives of tumor organoid[J]. China Oncology, 2024, 34(8): 763-776.

Figures/Tables 4

Fig. 1

Fig. 2

Tab. 1

Summary of organoid-guided interventional trials"

Identifiers	Histology	Registration year	Drug (s)	Status	Results/goals
-	Colorectal cancer^[33]	Unknown	A 87-drug panel	Completed	This trial enrolled 28 patients and successfully established 19 organoids, out of which 15 were utilized for drug screening. One patient underwent treatment based on the drug sensitivity results of the organoid and showed partial remission after 3 months of evaluation
2014-003811-13	Colorectal cancer^[34]	2014	Palbociclib, axitinib, selumetinib, gefitinib	Completed	This trial enrolled 54 patients and successfully established 31 organoids, of which 25 were utilized for drug screening. Out of the 25 organoids, 19 received drug sensitivity results. Based on these results, 6 patients underwent treatment; however, none of them showed any efficacy
NCT05842187	Pancreatic cancer/gastric cancer	2023	Various clinically approved drugs	Recruiting	This trial will use organoid models to guide the precision treatment of metastatic pancreatic cancer and gastric cancer
NCT04931394	Pancreatic cancer	2021	Gemcitabine, 5-fluorouracil, paclitaxel, oxaliplatin, irinotecan	Recruiting	This trial will use organoid models to guide the adjuvant chemotherapy of pancreatic cancer
NCT04931381	Pancreatic cancer	2021	Gemcitabine, 5-fluorouracil, paclitaxel, oxaliplatin, irinotecan	Recruiting	This trial will use organoid models to guide the chemotherapy of advanced pancreatic cancer
NCT06102824	Breast cancer	2023	Taxane, anthracycline, 5-fluorouracil, gemcitabine, vinorelbine, eribulin, utidelone, carboplatin, sacituzumab govitecan, and trastuzumab deruxtecan	Recruiting	This trial will use organoid models to guide the treatment of advanced breast cancer
NCT06268652	Breast cancer	2024	Personalized drug library contains 55 drugs approved by the FDA	Recruiting	This trial will compare the efficacy of organoid-guided personalized treatment with the treatment of physician’s choice in breast cancer
NCT05832398	Colorectal cancer	2023	Oxaliplatin, irinotecan, 5-fluorouracil	Recruiting	This trial will use organoid models to guide the precise chemotherapy of colorectal cancer
NCT05177432	Breast cancer	2021	10-12 anti-cancer drugs	Recruiting	This trial will use organoid models to develop a quadratic phenotypic optimization platform (QPOP) to guide the treatment of breast cancer
NCT05669586	Lung cancer	2023	Unknown	Recruiting	This trial will use organoid models to guide the precise treatment of refractory non-small cell lung cancer
NCT05813509	Ovarian cancer	2022	10 potential clinical therapeutic drugs	Recruiting	This trial will use organoid models to guide the personalized treatment of ovarian cancer
NCT06246630	Pancreatic neuroendocrine tumor	2024	Various clinically approved drugs	Not yet recruiting	This trial will use organoid models to guide the treatment of pancreatic neuroendocrine tumors
NCT05024734	Bladder cancer	2022	Epirubicin, mitomycin, gemcitabine, docetaxel	Recruiting	This trial will use organoid models to guide the treatment of non-muscle invasive bladder cancers
NCT06077591	Solid tumors	2024	Unknown	Not yet recruiting	This trial will use organoid models and next-generating sequencing to guide the treatment of advanced and inoperable solid tumors
NCT05352165	Colorectal cancer	2023	Oxaliplatin, irinotecan, 5-fluorouracil	Not yet recruiting	This trial will compare the efficacy of organoid-guided neoadjuvant chemotherapy with traditional neoadjuvant chemotherapy regimens in advanced colorectal cancer
NCT05378048	Abdominal tumors	2022	Unknown	Withdrawn	This trial will compare the efficacy of organoid-guided personalized treatment with traditional treatment strategies in advanced and inoperable abdominal tumors
NCT04279509	Solid tumors	2019	5-fluorouracil, carboplatin, cyclophosphamide, docetaxel, doxorubicin, gemcitabine, irinotecan, oxaliplatin, paclitaxel, vinorelbine, etoposide, ifosfamide, methotrexate, pemetrexed and topotecan	Unknown	This trial will use organoid models to conduct high-throughput drug screening to guide the chemotherapy of refractory solid tumors
NCT05267912	Any cancer type	2022	A panel of drugs (chemotherapy, hormonal therapy, targeted therapy)	Recruiting	This trial is a multi-center study evaluating the feasibility of using organoid models to guide the precision treatment of multiple advanced tumors
NCT04450706	Breast cancer	2021	Unknown	Recruiting	This trial will use organoid models to guide the precision treatment of metastatic breast cancers
NCT06057298	Colorectal cancer	2021	Oxaliplatin, mitomycin	Recruiting	This trial will use organoid models to guide the hyperthermic introperitoneal chemotherapy of colorectal cancers with peritoneal metastasis
NCT04842006	Colorectal cancer	2021	Capecitabine, oxaliplatin	Recruiting	This trial will use organoid models to guide the neoadjuvant and adjuvant therapy of colorectal cancer
NCT05725200	Colorectal cancer	2022	Alectinib, cetuximab, crizotinib, dasatinib, everolimus, encorafenib, gemcitabine, idelalisib, larotrectinib, methotrexate, palbociclib, panobinostat, pembrolizumab, petrozumab, trastuzumab, talazoparib, venetoclax	Recruiting	This trial will use organoid models to guide the precision treatment of metastatic colorectal cancer
NCT05464082	Breast cancer	2023	Unknown	Recruiting	This trial will use organoid models to guide the precision treatment of metastatic triple-negative breast cancers
NCT03778814	Lung cancer/solid tumors	2018	Engineering TCR-T cells	Recruiting	This trial will use organoid models to guide the TCR-T immunotherapy of lung cancers and other solid tumors
NCT05429684	Breast cancer	2021	Trastuzumab, pertuzumab, nab paclitaxel, pyrotinib, capecitabine, T-DM1, everolimus, CDK4/6 inhibitor, aromatase-Inhibitors, anti-PD-1 monoclonal antibody	Recruiting	This trial will use organoid models to guide the precision treatment of refractory HER2-positive breast cancers
NCT05381038	Solid tumors	2022	Azacitidine, docetaxel, paclitaxel, irinotecan	Not yet recruiting	This trial will use organoid models to develop a QPOP and CURATE.AI, which will guide the personalized combinatory therapy with azacytidine in solid tumors.
NCT06227065	Bladder cancer	2024	Epirubicin, mitomycin, gemcitabine, docetaxel	Not yet recruiting	This trial will use organoid models to guide the precise neoadjuvant chemotherapy of low-grade non-muscle invasive bladder cancers
NCT05432518	Glioblastoma	2023	Afatinib, dasatinib, palbociclib, everolimus, olaparib	Recruiting	This trial will use organoid models to guide the treatment of refractory glioblastoma
NCT05473923	Glioma	2022	Unknown	Recruiting	This trial will use organoid models to guide the precision treatment of refractory high-grade glioma
NCT05532397	Astrocytoma	2023	Unknown	Recruiting	This trial will use organoid models to guide the combinatory therapy of refractory high-grade astrocytoma
2020-003395-41	Colorectal cancer	2020	Alectinib, crizotinib, dasatinib, everolimus, gemcitabine, idelalisib, larotrectinib, methotrexate, palbociclib, panobinostat, pembrolizumab, pertuzumab, trastuzumab, talazoparib, venetoclax, cetuximab, encorafinib	Recruiting	This trial will use organoid models to guide the personalized therapy of colorectal cancers

Tab. 1

Fig. 3

References 68

[1]	SIEGEL R, GIAQUINTO A N, JEMAL A. Cancer statistics, 2024[J]. CA A Cancer J Clin, 2024, 74: 12-49.
[2]	郑荣寿, 陈茹, 韩冰峰, 等. 2022年中国恶性肿瘤流行情况分析[J]. 中华肿瘤杂志, 2024, 46(3):221-231.
	ZHENG R S, CHEN R, HAN B F, et al. Cancer incidence and mortality in China, 2022[J]. Chin J Oncol, 2024, 46(3): 221-231.
[3]	TSIMBERIDOU A M, FOUNTZILAS E, NIKANJAM M, et al. Review of precision cancer medicine: evolution of the treatment paradigm[J]. Cancer Treat Rev, 2020, 86: 102019.
[4]	XU H X, JIAO D C, LIU A G, et al. Tumor organoids: applications in cancer modeling and potentials in precision medicine[J]. J Hematol Oncol, 2022, 15(1): 58.
[5]	FOO M A, YOU M L, CHAN S L, et al. Clinical translation of patient-derived tumour organoids-bottlenecks and strategies[J]. Biomark Res, 2022, 10(1): 10.
[6]	VAN DE WETERING M, FRANCIES H E, FRANCIS J M, et al. Prospective derivation of a living organoid biobank of colorectal cancer patients[J]. Cell, 2015, 161(4): 933-945. doi: 10.1016/j.cell.2015.03.053 pmid: 25957691
[7]	PAULI C, HOPKINS B D, PRANDI D, et al. Personalized in vitro and in vivo cancer models to guide precision medicine[J]. Cancer Discov, 2017, 7(5): 462-477.
[8]	WILSON H V. A new method by which sponges may be artificially reared[J]. Science, 1907, 25(649): 912-915. pmid: 17842577
[9]	CORRÒ C, NOVELLASDEMUNT L, LI V S W. A brief history of organoids[J]. Am J Physiol Cell Physiol, 2020, 319(1): C151-C165.
[10]	EVANS M. Origin of mouse embryonal carcinoma cells and the possibility of their direct isolation into tissue culture[J]. J Reprod Fertil, 1981, 62(2): 625-631.
[11]	LI M L, AGGELER J, FARSON D A, et al. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells[J]. Proc Natl Acad Sci U S A, 1987, 84(1): 136-140.
[12]	SATO T, VRIES R G, SNIPPERT H J, et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche[J]. Nature, 2009, 459(7244): 262-265.
[13]	SATO T, STANGE D E, FERRANTE M, et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium[J]. Gastroenterology, 2011, 141(5): 1762-1772.
[14]	RAHRMANN E P, SHORTHOUSE D, JASSIM A, et al. The NALCN channel regulates metastasis and nonmalignant cell dissemination[J]. Nat Genet, 2022, 54(12): 1827-1838. doi: 10.1038/s41588-022-01182-0 pmid: 36175792
[15]	MAO Y N, WANG W, YANG J W, et al. Drug repurposing screening and mechanism analysis based on human colorectal cancer organoids[J]. Protein Cell, 2024, 15(4): 285-304. doi: 10.1093/procel/pwad038
[16]	KATO H, TATEISHI K, FUJIWARA H, et al. MNX1-HNF1B axis is indispensable for intraductal papillary mucinous neoplasm lineages[J]. Gastroenterology, 2022, 162(4): 1272-1287.e16.
[17]	ZHAO H, CHENG Y L, KALRA A, et al. Generation and multiomic profiling of a TP53/CDKN2A double-knockout gastroesophageal junction organoid model[J]. Sci Transl Med, 2022, 14(673): eabq6146.
[18]	DAYTON T L, ALCALA N, MOONEN L, et al. Druggable growth dependencies and tumor evolution analysis in patient-derived organoids of neuroendocrine neoplasms from multiple body sites[J]. Cancer Cell, 2023, 41(12): 2083-2099.e9. doi: 10.1016/j.ccell.2023.11.007 pmid: 38086335
[19]	GRIGER J, WIDHOLZ S A, JESINGHAUS M, et al. An integrated cellular and molecular model of gastric neuroendocrine cancer evolution highlights therapeutic targets[J]. Cancer Cell, 2023, 41(7): 1327-1344.e10.
[20]	LI Z C, XU H B, GONG Y Q, et al. Patient-derived upper tract urothelial carcinoma organoids as a platform for drug screening[J]. Adv Sci, 2022, 9(4): e2103999.
[21]	PONSIOEN B, POST J B, BUISSANT DES AMORIE J R, et al. Quantifying single-cell ERK dynamics in colorectal cancer organoids reveals EGFR as an amplifier of oncogenic MAPK pathway signalling[J]. Nat Cell Biol, 2021, 23(4): 377-390. doi: 10.1038/s41556-021-00654-5 pmid: 33795873
[22]	HERPERS B, EPPINK B, JAMES M I, et al. Functional patient-derived organoid screenings identify MCLA-158 as a therapeutic EGFR × LGR5 bispecific antibody with efficacy in epithelial tumors[J]. Nat Cancer, 2022, 3(4): 418-436.
[23]	ROOVERS R, HERPERS B, JAMES M, et al. Abstract 32: Preclinical evaluation of MCLA-158: a bispecific antibody targeting LGR5 and EGFR using patient-derived colon carcinoma organoids[J]. Cancer Res, 2017, 77(13_Supplement): 32.
[24]	COHEN E E, FAYETTE J, DASTE A, et al. Abstract CT012: Clinical activity of MCLA-158 (petosemtamab), an IgG1 bispecific antibody targeting EGFR and LGR5, in advanced head and neck squamous cell cancer (HNSCC)[J]. Cancer Res, 2023, 83(8_Suppl): CT012.
[25]	PAUL S M, MYTELKA D S, DUNWIDDIE C T, et al. How to improve R&D productivity: the pharmaceutical industry’s grand challenge[J]. Nat Rev Drug Discov, 2010, 9(3): 203-214.
[26]	ONOZATO D, YAMASHITA M, NAKANISHI A, et al. Generation of intestinal organoids suitable for pharmacokinetic studies from human induced pluripotent stem cells[J]. Drug Metab Dispos, 2018, 46(11): 1572-1580. doi: 10.1124/dmd.118.080374 pmid: 29615438
[27]	KATSUDA T, KAWAMATA M, HAGIWARA K, et al. Conversion of terminally committed hepatocytes to culturable bipotent progenitor cells with regenerative capacity[J]. Cell Stem Cell, 2017, 20(1): 41-55. doi: S1934-5909(16)30347-2 pmid: 27840021
[28]	VOGES H K, MILLS R J, ELLIOTT D A, et al. Development of a human cardiac organoid injury model reveals innate regenerative potential[J]. Development, 2017, 144(6): 1118-1127. doi: 10.1242/dev.143966 pmid: 28174241
[29]	TAKASATO M, ER P X, CHIU H S, et al. Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis[J]. Nature, 2016, 536(7615): 238.
[30]	国家药品监督管理局药品审评中心. 人源干细胞产品非临床研究技术指导原则[Z]. 2024.
	Center for Drug Evalution, National Medical Products Administration. Guidelines for non clinical research techniques of human stem cell products[Z]. 2024.
[31]	ZEHIR A, BENAYED R, SHAH R H, et al. Erratum: mutational landscape of metastatic cancer revealed from prospective clinical sequencing of 10 000 patients[J]. Nat Med, 2017, 23(8): 1004.
[32]	YAO Y, XU X Y, YANG L F, et al. Patient-derived organoids predict chemoradiation responses of locally advanced rectal cancer[J]. Cell Stem Cell, 2020, 26(1): 17-26.e6. doi: S1934-5909(19)30431-X pmid: 31761724
[33]	NARASIMHAN V, WRIGHT J A, CHURCHILL M, et al. Medium-throughput drug screening of patient-derived organoids from colorectal peritoneal metastases to direct personalized therapy[J]. Clin Cancer Res, 2020, 26(14): 3662-3670. doi: 10.1158/1078-0432.CCR-20-0073 pmid: 32376656
[34]	OOFT S N, WEEBER F, SCHIPPER L, et al. Prospective experimental treatment of colorectal cancer patients based on organoid drug responses[J]. ESMO Open, 2021, 6(3): 100103.
[35]	LI M, IZPISUA BELMONTE J C. Organoids-preclinical models of human disease[J]. N Engl J Med, 2019, 380(6): 569-579.
[36]	LESAVAGE B L, SUHAR R A, BROGUIERE N, et al. Next-generation cancer organoids[J]. Nat Mater, 2022, 21(2): 143-159.
[37]	VAN RENTERGHEM A W J, VAN DE HAAR J, VOEST E E. Functional precision oncology using patient-derived assays: bridging genotype and phenotype[J]. Nat Rev Clin Oncol, 2023, 20(5): 305-317. doi: 10.1038/s41571-023-00745-2 pmid: 36914745
[38]	WANG H, NING X F, ZHAO F, et al. Human organoids-on-chips for biomedical research and applications[J]. Theranostics, 2024, 14(2): 788-818. doi: 10.7150/thno.90492 pmid: 38169573
[39]	KIM M, PANAGIOTAKOPOULOU M, CHEN C, et al. Micro-engineering and nano-engineering approaches to investigate tumour ecosystems[J]. Nat Rev Cancer, 2023, 23(9): 581-599. doi: 10.1038/s41568-023-00593-3 pmid: 37353679
[40]	LIU H T, GAN Z Q, QIN X Y, et al. Advances in microfluidic technologies in organoid research[J]. Adv Healthc Mater, 2023: e2302686.
[41]	TAO T T, DENG P W, WANG Y Q, et al. Microengineered multi-organoid system from hiPSCs to recapitulate human liver-islet axis in normal and type 2 diabetes[J]. Adv Sci, 2022, 9(5): e2103495.
[42]	DORNHOF J, KIENINGER J, MURALIDHARAN H, et al. Microfluidic organ-on-chip system for multi-analyte monitoring of metabolites in 3D cell cultures[J]. Lab Chip, 2022, 22(2): 225-239.
[43]	YANG X Y, LI K Y, ZHANG X, et al. Nanofiber membrane supported lung-on-a-chip microdevice for anti-cancer drug testing[J]. Lab Chip, 2018, 18(3): 486-495. doi: 10.1039/c7lc01224a pmid: 29309077
[44]	AUNG A, KUMAR V, THEPRUNGSIRIKUL J, et al. An engineered tumor-on-a-chip device with breast cancer-immune cell interactions for assessing T-cell recruitment[J]. Cancer Res, 2020, 80(2): 263-275. doi: 10.1158/0008-5472.CAN-19-0342 pmid: 31744818
[45]	PARK D, SON K, HWANG Y, et al. High-throughput microfluidic 3D cytotoxicity assay for cancer immunotherapy (CACI-IMPACT platform)[J]. Front Immunol, 2019, 10: 1133. doi: 10.3389/fimmu.2019.01133 pmid: 31191524
[46]	TANG X Y, WU S S, WANG D, et al. Human organoids in basic research and clinical applications[J]. Signal Transduct Target Ther, 2022, 7(1): 168.
[47]	CAKIR B, XIANG Y F, TANAKA Y, et al. Engineering of human brain organoids with a functional vascular-like system[J]. Nat Methods, 2019, 16(11): 1169-1175. doi: 10.1038/s41592-019-0586-5 pmid: 31591580
[48]	SHIRURE V S, BI Y, CURTIS M B, et al. Tumor-on-a-chip platform to investigate progression and drug sensitivity in cell lines and patient-derived organoids[J]. Lab Chip, 2018, 18(23): 3687-3702. doi: 10.1039/c8lc00596f pmid: 30393802
[49]	LIU H T, ZHANG X, LIU J Y, et al. Vascularization of engineered organoids[J]. BMEMat, 2023, 1(3): e12031.
[50]	RAJASEKAR S, LIN D S Y, ABDUL L, et al. IFlowPlate-a customized 384-well plate for the culture of perfusable vascularized colon organoids[J]. Adv Mater, 2020, 32(46): e2002974.
[51]	PUCA L, BAREJA R, PRANDI D, et al. Patient derived organoids to model rare prostate cancer phenotypes[J]. Nat Commun, 2018, 9(1): 2404.
[52]	KARKAMPOUNA S, MANNA F L, BENJAK A, et al. Patient-derived xenografts and organoids model therapy response in prostate cancer[J]. Nat Commun, 2021, 12(1): 1117.
[53]	LANDON-BRACE N, LI N T, MCGUIGAN A P. Exploring new dimensions of tumor heterogeneity: the application of single cell analysis to organoid-based 3D in vitro models[J]. Adv Healthc Mater, 2023, 12(26): e2300903.
[54]	HUANG L X, XU Y Q, WANG N, et al. Next-generation preclinical functional testing models in cancer precision medicine: CTC-derived organoids[J]. Small Methods, 2024, 8(1): e2301009.
[55]	KINOSHITA K, TSUKAMOTO Y, HIRASHITA Y, et al. Efficient establishment of bile-derived organoids from biliary cancer patients[J]. Lab Invest, 2023, 103(6): 100105.
[56]	TUVESON D, CLEVERS H. Cancer modeling meets human organoid technology[J]. Science, 2019, 364(6444): 952-955. doi: 10.1126/science.aaw6985 pmid: 31171691
[57]	AISENBREY E A, MURPHY W L. Synthetic alternatives to Matrigel[J]. Nat Rev Mater, 2020, 5(7): 539-551. doi: 10.1038/s41578-020-0199-8 pmid: 32953138
[58]	CLINTON J, MCWILLIAMS-KOEPPEN P. Initiation, expansion, and cryopreservation of human primary tissue-derived normal and diseased organoids in embedded three-dimensional culture[J]. Curr Protoc Cell Biol, 2019, 82(1): e66.
[59]	DAO V, YUKI K, LO Y H, et al. Immune organoids: from tumor modeling to precision oncology[J]. Trends Cancer, 2022, 8(10): 870-880. doi: 10.1016/j.trecan.2022.06.001 pmid: 35773148
[60]	VOABIL P, BRUIJN M D, ROELOFSEN L M, et al. An ex vivo tumor fragment platform to dissect response to PD-1 blockade in cancer[J]. Nat Med, 2021, 27(7): 1250-1261. doi: 10.1038/s41591-021-01398-3 pmid: 34239134
[61]	国家卫生健康委, 教育部, 科技部, 等. 涉及人的生命科学和医学研究伦理审查办法[Z]. 2023.
	National Health Commission, Ministry of Education, National Health Commission. et al. Methods for ethical review of human life sciences and medical research[Z]. 2023.
[62]	类器官药物敏感性检测指导肿瘤精准治疗临床应用专家共识(2022年版)编写专家组. 类器官药物敏感性检测指导肿瘤精准治疗临床应用专家共识(2022年版)[J]. 中国癌症防治杂志, 2022, 14(3): 234-239.
	Expert Consensus on the Clinical Application of Precision Cancer Treatment Guided by Sensitivity Testing of Organoids (2022 Edition) Writing Expert Group. Expert consensus on clinical application of drug sensitivity test of organ-like drugs to guide accurate treatment of tumors (2022 edition)[J]. Chin J Oncol Prev Treat, 2022, 14(3): 234-239.
[63]	NIIKAWA T, HAYASHI Y, SHEPHERD J, et al. Human brain organoids and consciousness[J]. Neuroethics, 2022, 15(1): 5.
[64]	JONGH D D, MASSEY E K, VANGUARD CONSORTIUM, et al. Organoids: a systematic review of ethical issues[J]. Stem Cell Res Ther, 2022, 13(1): 337.
[65]	Bold Goals for U.S. Biotechnology and Biomanufacturing[Z]. 2023.
[66]	Horizon Europe Work Programme 2023-2024[Z]. 2023.
[67]	科技部基础研究司. “十四五”国家重点研发计划“干细胞研究与器官修复”重点专项2021年度项目申报指南[Z]. 2021.
	Department of Basic Research, Ministry of Science and Technology of People’s Republic of China. Guidelines for the application of key special projects for stem cell research and organ repair under the 14th Five Year Plan of China in 2021[Z]. 2021.
[68]	科技部基础研究司. “十四五”国家重点研发计划“干细胞研究与器官修复”重点专项2022年度项目申报指南[Z]. 2022.
	Department of Basic Research, Ministry of Science and Technology of People’s Republic of China. Guidelines for the application of key special projects for stem cell research and organ repair under the 14th Five Year Plan of China in 2022[Z]. 2022.