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

作者：Bei, Yang; Yeung, Enoch; Lui, Cecillia; Ong, Chin Siang; Pitaktong, Isaree; Huang, Chenyu; Inoue, Takahiro; Matsushita, Hiroshi; Ma, Chunye; Hibino, Narutoshi^*

来源：Jove-Journal of Visualized Experiments, 2018, (138): e58252.

DOI：10.3791/58252

• Bioengineering
• Issue 138
• Cardiac tissue engineering
• hanging drop
• spheroids
• net-mold
• scaffold-free
• heart failure

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.

• 出版日期2018-8
• 单位吉林大学