Myocardial Tissue Engineering With Cells Derived From Human-Induced Pluripotent Stem Cells and a Native-Like, High-Resolution, 3-Dimensionally Printed Scaffold.
Myocardial Tissue Engineering With Cells Derived From Human-Induced Pluripotent Stem Cells and a Native-Like, High-Resolution, 3-Dimensionally Printed Scaffold.
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DOI：
10.1161/circresaha.116.310277
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发表时间：
2017-04-14
影响因子：
20.1
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Gao L;Kupfer ME;Jung JP;Yang L;Zhang P;Da Sie Y;Tran Q;Ajeti V;Freeman BT;Fast VG;Campagnola PJ;Ogle BM;Zhang J
传统的三维（3D）打印技术不能产生单个细胞相互作用的大小的结构。在这里，我们使用多光子激发的三维打印（MPE-3DP）来生成一个亚微米分辨率的天然样细胞外基质（ECM）支架，然后将从人诱导多能干细胞（iPSCs）分化出来的心肌细胞（CMs）、平滑肌细胞（SMCs）和内皮细胞（ECs）植入支架，生成一个人ipsc衍生的心肌贴片（hCMP），随后在小鼠心肌梗死（MI）模型中进行评估。将约50,000个人，ipsc衍生的CMs， SMCs和ECs（以2:1:1的比例）植入支架以产生hCMP， hCMP在植入后1天内开始产生钙瞬态并同步跳动；在接下来的7天内，收缩和舒张的速度以及钙瞬态的峰值幅度显著增加。在手术诱导心肌梗死小鼠中进行测试时，治疗后第4周，hcmp处理动物的心功能、梗死面积、凋亡、血管和小动脉密度以及细胞增殖的测量结果明显优于无细胞支架处理动物，hcmp处理动物的细胞植入率在第1周为24.5%，第4周为11.2%。因此，新的MPE-3DP技术产生的基于ecm的支架具有优异的分辨率和保真度，用这些支架制造的hcmp可以显著提高缺血性心肌损伤的恢复。
Conventional three-dimensional (3D) printing techniques cannot produce structures of the size at which individual cells interact. Here, we used multiphoton-excited, 3-dimensional printing (MPE-3DP) to generate a native-like, extracellular matrix (ECM) scaffold with submicron resolution, and then seeded the scaffold with cardiomyocytes (CMs), smooth-muscle cells (SMCs), and endothelial cells (ECs) that had been differentiated from human induced-pluripotent stem cells (iPSCs) to generate a human, iPSC-derived cardiac muscle patch (hCMP), which was subsequently evaluated in a murine model of myocardial infarction (MI). The scaffold was seeded with ~50,000 human, iPSC-derived CMs, SMCs, and ECs (in a 2:1:1 ratio) to generate the hCMP, which began generating calcium transients and beating synchronously within 1 day of seeding; the speeds of contraction and relaxation and the peak amplitudes of the calcium transients increased significantly over the next 7 days. When tested in mice with surgically induced MI, measurements of cardiac function, infarct size, apoptosis, both vascular and arteriole density, and cell proliferation at week 4 after treatment were significantly better in animals treated with the hCMPs than in animals treated with cell-free scaffolds, and the rate of cell engraftment in hCMP-treated animals was 24.5% at week 1 and 11.2% at week 4. Thus, the novel MPE-3DP technique produces ECM-based scaffolds with exceptional resolution and fidelity, and hCMPs fabricated with these scaffolds may significantly improve recovery from ischemic myocardial injury.