Ke Ming, Xu Wenhui, Hao Yansha, Zheng Feiyang, Yang Guanyuan, Fan Yonghong, Wang Fangfang, Nie Zhiqiang, Zhu Chuhong
Department of Anatomy, Third Military Medical University, Chongqing 400038, China.
State Key Laboratory of Trauma, Burn and Combined Injury, Chongqing 400038, China.
Regen Biomater. 2023 Dec 29;11:rbad117. doi: 10.1093/rb/rbad117. eCollection 2024.
Engineering myocardium has shown great clinal potential for repairing permanent myocardial injury. However, the lack of perfusing blood vessels and difficulties in preparing a thick-engineered myocardium result in its limited clinical use. We prepared a mixed gel containing fibrin (5 mg/ml) and collagen I (0.2 mg/ml) and verified that human umbilical vein endothelial cells (HUVECs) and human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) could form microvascular lumens and myocardial cell clusters by harnessing the low-hardness and hyperelastic characteristics of fibrin. hiPSC-CMs and HUVECs in the mixed gel formed self-organized cell clusters, which were then cultured in different media using a three-phase approach. The successfully constructed vascularized engineered myocardial tissue had a spherical structure and final diameter of 1-2 mm. The tissue exhibited autonomous beats that occurred at a frequency similar to a normal human heart rate. The internal microvascular lumen could be maintained for 6 weeks and showed good results during preliminary surface re-vascularization and vascular remodeling . In summary, we propose a simple method for constructing vascularized engineered myocardial tissue, through phased cultivation that does not rely on high-end manufacturing equipment and cutting-edge preparation techniques. The constructed tissue has potential value for clinical use after preliminary evaluation.
工程化心肌在修复永久性心肌损伤方面已显示出巨大的临床潜力。然而，缺乏灌注血管以及制备厚层工程化心肌的困难导致其临床应用受限。我们制备了一种含有纤维蛋白（5毫克/毫升）和I型胶原蛋白（0.2毫克/毫升）的混合凝胶，并证实人脐静脉内皮细胞（HUVECs）和人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）可以利用纤维蛋白的低硬度和超弹性特性形成微血管腔和心肌细胞簇。混合凝胶中的hiPSC-CMs和HUVECs形成了自组织细胞簇，然后采用三相方法在不同培养基中培养。成功构建的血管化工程化心肌组织具有球形结构，最终直径为1-2毫米。该组织表现出自主搏动，搏动频率与正常人类心率相似。内部微血管腔可维持6周，在初步表面再血管化和血管重塑过程中显示出良好效果。总之，我们提出了一种构建血管化工程化心肌组织的简单方法，通过分阶段培养，该方法不依赖高端制造设备和前沿制备技术。经初步评估，构建的组织具有临床应用的潜在价值。
