Construction of cardiac organoids derived from human induced pluripotent stem cells for cardiac disease modeling and drug evaluation.
10.12122/j.issn.1673-4254.2025.11.17
- Author:
Xue GONG
1
;
Yongyang FAN
2
;
Kaiyuan LUO
2
;
Yi YAN
2
;
Zhonghao LI
2
Author Information
1. General Hospital of Southern Theater Command of PLA, Guangzhou 510010, China.
2. Department of Cardiology, Third Affiliated Hospital of Guangzhou Medical University//Guangdong Provincial Key Laboratory of Major Obstetric Diseases//Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, Guangzhou 510150, China.
- Publication Type:Journal Article
- Keywords:
cardiac organoids;
disease modeling;
drug evaluation;
human induced pluripotent stem cells
- MeSH:
Humans;
Induced Pluripotent Stem Cells/cytology*;
Organoids/cytology*;
Myocytes, Cardiac/cytology*;
Cell Differentiation;
Heart Diseases
- From:
Journal of Southern Medical University
2025;45(11):2444-2455
- CountryChina
- Language:Chinese
-
Abstract:
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.
CONCLUSIONS:Cardiac organoids derived from human induced pluripotent stem cells have been successfully constructed and can be used for cardiac disease modeling and drug evaluation.
