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J Vis Exp. 2018 Aug 5;(138). doi: 10.3791/58252.

A Net Mold-based Method of Scaffold-free Three-Dimensional Cardiac Tissue Creation.

Author information

1
Department of Cardiac Surgery, The First Hospital of Jilin University; Division of Cardiac Surgery, Johns Hopkins Hospital.
2
Division of Cardiac Surgery, Johns Hopkins Hospital.
3
Division of Cardiac Surgery, Johns Hopkins Hospital; nhibino1@jhmi.edu.

Abstract

This protocol describes a novel and easy net mold-based method to create three-dimensional (3-D) cardiac tissues without additional scaffold material. Human-induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs), human cardiac fibroblasts (HCFs), and human umbilical vein endothelial cells (HUVECs) are isolated and used to generate a cell suspension with 70% iPSC-CMs, 15% HCFs, and 15% HUVECs. They are co-cultured in an ultra-low attachment "hanging drop" system, which contains micropores for condensing hundreds of spheroids at one time. The cells aggregate and spontaneously form beating spheroids after 3 days of co-culture. The spheroids are harvested, seeded into a novel mold cavity, and cultured on a shaker in the incubator. The spheroids become a mature functional tissue approximately 7 days after seeding. The resultant multilayered tissues consist of fused spheroids with satisfactory structural integrity and synchronous beating behavior. This new method has promising potential as a reproducible and cost-effective method to create engineered tissues for the treatment of heart failure in the future.

PMID:
30124650
PMCID:
PMC6126624
[Available on 2020-08-05]
DOI:
10.3791/58252
[Indexed for MEDLINE]

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