A net mold-based method of scaffold-free three-dimensional cardiac tissue creation

Yang Bai, Enoch Yeung, Cecillia Lui, Chin Siang Ong, Isaree Pitaktong, Chenyu Huang, Takahiro Inoue, Hiroshi Matsushita, Chunye Ma, Narutoshi Hibino

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


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.

Original languageEnglish (US)
Article numbere58252
JournalJournal of Visualized Experiments
Issue number138
StatePublished - Aug 5 2018


  • Bioengineering
  • Cardiac tissue engineering
  • Hanging drop
  • Heart failure
  • Issue 138
  • Net-mold
  • Scaffold-free
  • Spheroids

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology


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