학술논문
Generation, functional analysis and applications of isogenic three-dimensional self-aggregating cardiac microtissues from human pluripotent stem cells
Document Type
Report
Author
Source
Nature Protocols. April 2021, Vol. 16 Issue 4, p2213, 44 p.
Subject
Language
English
ISSN
1754-2189
Abstract
Author(s): Giulia Campostrini [sup.1] , Viviana Meraviglia [sup.1] , Elisa Giacomelli [sup.1] [sup.5] , Ruben W. J. van Helden [sup.1] , Loukia Yiangou [sup.1] , Richard P. Davis [sup.1] , [...]
Tissue-like structures from human pluripotent stem cells containing multiple cell types are transforming our ability to model and understand human development and disease. Here we describe a protocol to generate cardiomyocytes (CMs), cardiac fibroblasts (CFs) and cardiac endothelial cells (ECs), the three principal cell types in the heart, from human induced pluripotent stem cells (hiPSCs) and combine them in three-dimensional (3D) cardiac microtissues (MTs). We include details of how to differentiate, isolate, cryopreserve and thaw the component cells and how to construct and analyze the MTs. The protocol supports hiPSC-CM maturation and allows replacement of one or more of the three heart cell types in the MTs with isogenic variants bearing disease mutations. Differentiation of each cell type takes ~30 d, while MT formation and maturation requires another 20 d. No specialist equipment is needed and the method is inexpensive, requiring just 5,000 cells per MT. Human induced pluripotent stem cells are differentiated to the three main cell types found in the heart--cardiomyocytes, cardiac fibroblasts and cardiac endothelial cells--and then combined to form 3D cardiac microtissues.
Tissue-like structures from human pluripotent stem cells containing multiple cell types are transforming our ability to model and understand human development and disease. Here we describe a protocol to generate cardiomyocytes (CMs), cardiac fibroblasts (CFs) and cardiac endothelial cells (ECs), the three principal cell types in the heart, from human induced pluripotent stem cells (hiPSCs) and combine them in three-dimensional (3D) cardiac microtissues (MTs). We include details of how to differentiate, isolate, cryopreserve and thaw the component cells and how to construct and analyze the MTs. The protocol supports hiPSC-CM maturation and allows replacement of one or more of the three heart cell types in the MTs with isogenic variants bearing disease mutations. Differentiation of each cell type takes ~30 d, while MT formation and maturation requires another 20 d. No specialist equipment is needed and the method is inexpensive, requiring just 5,000 cells per MT. Human induced pluripotent stem cells are differentiated to the three main cell types found in the heart--cardiomyocytes, cardiac fibroblasts and cardiac endothelial cells--and then combined to form 3D cardiac microtissues.