南方医科大学学报2025，Vol.45Issue(11)：2444-2455,12.DOI:10.12122/j.issn.1673-4254.2025.11.17
利用人诱导多能干细胞构建的心脏类器官在心脏疾病建模及药物评价中的应用价值
Construction of cardiac organoids derived from human induced pluripotent stem cells for cardiac disease modeling and drug evaluation
摘要
Abstract
Objective To explore the potential applications of human induced pluripotent stem cell-derived cardiac organoids in constructing cardiac injury models and drug evaluation.Methods Cardiac organoids derived from the self-assembled human induced pluripotent stem cells were constructed by regulating the Wnt signaling pathway.Flow cytometry was used to detect the proportion of cardiomyocytes in the cardiac organoids,and RT-qPCR was employed to detect the mRNA expressions.Immunofluorescence staining was used to detect the protein expressions of TNNT2,CD31,and vimentin.The beating amplitude of the cardiac organoids was determined with calcium transient.In vitro myocardial injury models and ischemia-reperfusion models were established,and the cell injuries were examined using Masson staining.TUNEL staining and calcium transient detection were used to evaluate the adverse effects of doxorubicin and trastuzumab in the cardiac organoids.Results The cardiac organoids began to beat on the 8th day of culture and consisted of 32.4%cardiomyocytes with high expressions of the myocardial markers TNNT2,NKX2.5,RYR2 and KCNJ2.No significant differences in morphological size,beating frequency,proportion of cardiomyocytes,or myocardial contractility were observed in the cardiac organoids differentiated from different batches.These cardiac organoids could be maintained in in vitro culture conditions for at least 50 days.Captopril treatment could obviously alleviate liquid nitrogen-induced myocardial injury in the cardiac organoids.Hypoxia/reoxygenation induced ischemia-reperfusion injury and promoted myocardial fibrosis and apoptosis in the cardiac organoids.Treatment with doxorubicin for 24 h resulted in significantly increased cell death and reduced beating frequency and cell viability in the cardiac organoids in a dose-dependent manner.Trastuzumab significantly impaired the contractile and calcium handling abilities of the cardiac organoids.Conclusion Cardiac organoids derived from human induced pluripotent stem cells have been successfully constructed and can be used for cardiac disease modeling and drug evaluation.关键词
人诱导多能干细胞/心脏类器官/疾病建模/药物评价Key words
human induced pluripotent stem cells/cardiac organoids/disease modeling/drug evaluation引用本文复制引用
龚雪,樊雍扬,罗开元,燕翼,李忠豪..利用人诱导多能干细胞构建的心脏类器官在心脏疾病建模及药物评价中的应用价值[J].南方医科大学学报,2025,45(11):2444-2455,12.基金项目
国家自然科学基金(82570445) （82570445）
广东省科技厅国际合作专项(2022A0505050079) （2022A0505050079）
广东省基础与应用基础研究基金面上项目(2024A1515012365) （2024A1515012365）
广州市科技局临床研究专项(202201020189) Supported by National Natural Science Foundation of China(82570445). （202201020189）
