Establishment of 3D organoids model of cardiac hypertrophy and its application in the mechanistic analysis of cardiovascular traditional Chinese medicine
10.16438/j.0513-4870.2021-1756
- VernacularTitle:3D类器官心脏肥大模型的建立及在心血管病治疗中药作用机制解析中的应用
- Author:
Si-wen FAN
1
;
Yu-han ZHAO
1
;
Guang-xu XIAO
1
;
Guan-wei FAN
2
;
Yan ZHU
1
Author Information
1. State Key Laboratory of Component-based Chinese Medicine of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Chinese Medicine New Drug Research and Development Center, International Biomedical Research Institute, Tianjin 300457, China
2. State Key Laboratory of Component-based Chinese Medicine of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Tianjin Key Laboratory of Translational Research of TCM Prescription and Syndrome, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
- Publication Type:Research Article
- Keywords:
organoid;
cardiac microsphere;
cardiac hypertrophy;
italic>in vitro model;
traditional Chinese medicine;
traditional Chinese medicine injection;
high content imaging
- From:
Acta Pharmaceutica Sinica
2022;57(10):3067-3076
- CountryChina
- Language:Chinese
-
Abstract:
Compared with the traditional two-dimensional (2D) monolayer culture, three-dimensional (3D) organoid can better simulate the physiological and pathological microenvironment of organs and tissues. In this study, 3D cardiac organoids were constructed using cardiac fibroblasts (CFs), cardiac myocytes (CMs) and endothelial cells (ECs) isolated from hearts of 1-3-day Sprague-Dawley (SD) neonatal rats. The experimental scheme was approved by the Experimental Animal Welfare and Ethics Committee of Tianjin University of Traditional Chinese Medicine and met the standards of experimental animal welfare and ethics. Optimal seeding cell density and culture time were determined by observing the sphere diameter and pulsation. The hierarchical structure and cardiac-like function were evaluated by fluorescence staining. The results showed that the cardiac-like microspheres constructed with cell number of 1×104 still beated spontaneously even after 34 days in culture, and maintained characteristic cellular hierarchical structure. Then, based on these cardiac microspheres, a phenylephrine (PE)-induced cardiac hypertrophy model was established and evaluated by mitochondrial mass, intracellular Ca2+ concentration and mitochondrial membrane potential. Guanxinning Injection (GXNI) was tested to verify that the established model can be used for myocardial hypertrophy drug screen. The results showed that GXNI significantly reversed the enlargement of cardiac microsphere area and diameter, the increase of mitochondrial mass, intracellular Ca2+ concentration and the decrease of mitochondrial membrane potential caused by PE, and reduced upregulation of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC). In conclusion, this study successfully established a 3D in vitro model of cardiac remodeling induced by cardiac hypertrophy. In this new system, cardiac microspheres not only have cardiac-like morphology and extracellular matrix components, but also exhibit spontaneous and rhythmic systolic and diastolic function. Therefore, the cardiac microsphere is an effective model to investigate the pathological mechanism of cardiac hypertrophy and screen related drugs.
