Generation, functional analysis and applications of isogenic three-dimensional self-aggregating cardiac microtissues from human pluripotent stem cells.

Generation, functional analysis and applications of isogenic three-dimensional self-aggregating cardiac microtissues from human pluripotent stem cells.
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DOI:
10.1038/s41596-021-00497-2
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发表时间:
2021-04
期刊:
影响因子:
14.8
通讯作者:
Mummery CL
Mummery CL
中科院分区:
生物学1区
文献类型:
--
作者:
Campostrini G;Meraviglia V;Giacomelli E;van Helden RWJ;Yiangou L;Davis RP;Bellin M;Orlova VV;Mummery CL

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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 and cardiac endothelial cells, the three principal cell types in the heart, from human induced pluripotent stem cells (hiPSCs) and combine these in three-dimensional 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 approximately 30 days, while MT formation and maturation requires another 20 days. No specialist equipment is needed and the method is low cost, requiring just 5,000 cells per MT.
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