Gao Ling, Kupfer Molly E, Jung Jangwook P, Yang Libang, Zhang Patrick, Da Sie Yong, Tran Quyen, Ajeti Visar, Freeman Brian T, Fast Vladimir G, Campagnola Paul J, Ogle Brenda M, Zhang Jianyi
From the Department of Biomedical Engineering, School of Medicine, School of Engineering, University of Alabama at Birmingham (L.G., V.G.F., J.Z.); Department of Biomedical Engineering, University of Minnesota, Twin Cities, Minneapolis (M.E.K., J.P.J., L.Y., P.Z., B.T.F., B.M.O.); and Department of Biomedical Engineering, University of Wisconsin, Madison (Y.D.S., Q.T., V.A., P.J.C.).
Circ Res. 2017 Apr 14;120(8):1318-1325. doi: 10.1161/CIRCRESAHA.116.310277. Epub 2017 Jan 9.
Conventional 3-dimensional (3D) printing techniques cannot produce structures of the size at which individual cells interact.
Here, we used multiphoton-excited 3D printing to generate a native-like extracellular matrix scaffold with submicron resolution and then seeded the scaffold with cardiomyocytes, smooth muscle cells, and endothelial cells that had been differentiated from human-induced pluripotent stem cells to generate a human-induced pluripotent stem cell-derived cardiac muscle patch (hCMP), which was subsequently evaluated in a murine model of myocardial infarction.
The scaffold was seeded with ≈50 000 human-induced pluripotent stem cell-derived cardiomyocytes, smooth muscle cells, and endothelial cells (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 myocardial infarction, 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 multiphoton-excited 3D printing technique produces extracellular matrix-based scaffolds with exceptional resolution and fidelity, and hCMPs fabricated with these scaffolds may significantly improve recovery from ischemic myocardial injury.
传统的三维（3D）打印技术无法制造出单个细胞相互作用大小的结构。
在此，我们使用多光子激发3D打印技术生成具有亚微米分辨率的天然细胞外基质支架，然后将从人诱导多能干细胞分化而来的心肌细胞、平滑肌细胞和内皮细胞接种到该支架上，以生成人诱导多能干细胞衍生的心肌补片（hCMP），随后在心肌梗死小鼠模型中对其进行评估。
将约50000个人诱导多能干细胞衍生的心肌细胞、平滑肌细胞和内皮细胞（比例为2:1:1）接种到支架上以生成hCMP，接种后1天内hCMP开始产生钙瞬变并同步跳动；在接下来的7天里，收缩和舒张速度以及钙瞬变的峰值幅度显著增加。在手术诱导心肌梗死的小鼠中进行测试时，治疗后第4周，接受hCMP治疗的动物在心脏功能、梗死面积、细胞凋亡、血管和小动脉密度以及细胞增殖方面的测量结果均显著优于接受无细胞支架治疗的动物，hCMP治疗动物的细胞植入率在第1周为24.5%，第4周为11.2%。
因此，新型多光子激发3D打印技术可制造出具有卓越分辨率和保真度的基于细胞外基质的支架，用这些支架制造的hCMP可能显著改善缺血性心肌损伤后的恢复情况。
