基于hiPSC-CM的新型明胶水凝胶构建体外心肌微组织研究

Study on construction of myocardial microtissue in vitro by novel gelatin hydrogel combined with human induced pluripotent stem cell derived cardiomyocytes

目的 探索应用新型明胶水凝胶复合人诱导多能干细胞来源心肌细胞(hiPSC-CMs)构建体外心肌微组织的可行性。方法 制备一种新型明胶水凝胶,以冷冻扫描电镜观察其表面结构,以循环拉伸试验测定其机械强度。在该材料上种植hiPSC-CMs,评估该材料的细胞相容性及其沟槽结构对心肌细胞的影响。将交联明胶水凝胶与hiPSC-CMs混合制备心肌微组织,探索构建体外心肌微组织过程中和冻存后心肌细胞的活性。结果 冷冻扫描电镜显示,新型明胶水凝胶表面呈现均匀的疏松多孔结构。循环拉伸试验显示,新型明胶水凝胶具有良好的机械性能。细胞试验显示,心肌细胞在该材料上生长良好,其沟槽结构对心肌细胞的有序排列有一定的引导作用。新型明胶水凝胶与hiPSC-CMs混合构建心肌微组织的实验结果表明,明胶水凝胶对心肌细胞具有良好的保护作用,对冻存复苏的心肌细胞具有一定的细胞保护作用。结论 新型明胶水凝胶具有疏松多孔的表面微结构、良好的力学性能和细胞相容性,对体外构建心肌微组织中的细胞和冻存后的细胞具有保护作用,未来有望用于组织工程领域,进行器官再生、药物筛选、体外建模等方面的研究。

Objective To explore the feasibility of a gelatin hydrogel combined with human-induced pluripotent stem cells derived cardiomyocytes(hiPSC-CMs) to construct myocardial microtissue in vitro. Methods A novel gelatin hydrogel was prepared, and its surface structure was observed by freeze-scanning electron microscope and its mechanical strength was measured by cyclic tensile test. HiPSC-CMs were implanted into the materials to evaluate their compatibility and the effect of groove structure on cardiomyocytes. Futhermore, cross-linked gelatin hydrogel was mixed with hiPSC-CMs for myocardial microtissue to explore the viability of cardiomyocytes during the construction of myocardial microtissue in vitro and after freezing. Results The surface of the gelatin hydrogel showed uniform porous structure by freeze-scanning electron microscopy. The cyclic tensile test showed that the cross-linked gelatin hydrogels had good mechanical properties. The cell test showed that the cardiomyocytes grew well on the materials, and the channel hydrogel could guide the orderly arrangement of the cardiomyocytes. Gelatin hydrogel was mixed with hiPSC-CMs for myocardial microtissue, the results showed that the gelatin hydrogel had a good protective effect on cardiomyocytes during myocardial microtissue construction, and had some protective effects on the frozen-thawed and resuscitated cardiomyocytes. Conclusion The novel gelatin hydrogel has porous surface microstructure, good mechanical properties and cell compatibility. It can protect cells in the process of myocardial microtissue and after cryopreservation. It is hopeful to be used in tissue engineering in the future for organ regeneration, drug screening and in vitro modeling.