源自人诱导多能干细胞的网状工程化心脏组织用于体内心肌修复的制备

Preparation of Mesh-Shaped Engineered Cardiac Tissues Derived from Human iPS Cells for In Vivo Myocardial Repair.

作者信息

Nakane Takeichiro, Abulaiti Mosha, Sasaki Yuko, Kowalski William J, Keller Bradley B, Masumoto Hidetoshi

机构信息

Clinical Translational Research Program, RIKEN Center for Biosystems Dynamics Research; Department of Cardiovascular Surgery, Graduate School of Medicine, Kyoto University; Department of Cardiovascular Surgery, Mitsubishi Kyoto Hospital.

Clinical Translational Research Program, RIKEN Center for Biosystems Dynamics Research; Department of Cardiovascular Surgery, Graduate School of Medicine, Kyoto University.

出版信息

J Vis Exp. 2020 Jun 9(160). doi: 10.3791/61246.

DOI:10.3791/61246

PMID:32597877

Abstract

The current protocol describes methods to generate scalable, mesh-shaped engineered cardiac tissues (ECTs) composed of cardiovascular cells derived from human induced pluripotent stem cells (hiPSCs), which are developed towards the goal of clinical use. HiPSC-derived cardiomyocytes, endothelial cells, and vascular mural cells are mixed with gel matrix and then poured into a polydimethylsiloxane (PDMS) tissue mold with rectangular internal staggered posts. By culture day 14 ECTs mature into a 1.5 cm x 1.5 cm mesh structure with 0.5 mm diameter myofiber bundles. Cardiomyocytes align to the long-axis of each bundle and spontaneously beat synchronously. This approach can be scaled up to a larger (3.0 cm x 3.0 cm) mesh ECT while preserving construct maturation and function. Thus, mesh-shaped ECTs generated from hiPSC-derived cardiac cells may be feasible for cardiac regeneration paradigms.

摘要

当前方案描述了生成由源自人诱导多能干细胞（hiPSC）的心血管细胞组成的可扩展、网状工程化心肌组织（ECT）的方法，这些方法是朝着临床应用的目标开发的。将源自hiPSC的心肌细胞、内皮细胞和血管壁细胞与凝胶基质混合，然后倒入具有矩形内部交错柱的聚二甲基硅氧烷（PDMS）组织模具中。到培养第14天时，ECT成熟为1.5厘米×1.5厘米的网状结构，具有直径0.5毫米的肌纤维束。心肌细胞与每个束的长轴对齐并自发同步跳动。这种方法可以扩大到更大的（3.0厘米×3.0厘米）网状ECT，同时保持构建体的成熟和功能。因此，由源自hiPSC的心脏细胞生成的网状ECT可能适用于心脏再生范例。