Construction of multi-layered cardiomyocyte sheets using magnetite nanoparticles and magnetic force
Construction of multi-layered cardiomyocyte sheets using magnetite nanoparticles and magnetic force
复制标题
DOI：
10.1002/bit.21094
复制
发表时间：
2007-03-01
影响因子：
3.8
通讯作者：
Honda, Hiroyuki
中科院分区：
文献类型：
--
作者：
Shimizu, Kazunori;Ito, Akira;Honda, Hiroyuki
心脏组织工程需要构建由心肌细胞（CM）组成的三维（3-D）组织，这些心肌细胞彼此紧密连接。本研究的目的是使用基于磁力的组织工程（Mag-TE）构建“无支架”的多层3-D CM片材，并评估CM片材内细胞与细胞的功能连接。从2日龄Wistar大鼠分离的CM吸收具有正表面电荷（有利于吸附至靶细胞表面）的原始磁铁矿阳离子脂质体（MCL）。当将MCL以25、50和100 pg/细胞的磁铁矿浓度加入CM培养基中时，随后的测量结果显示，在37 ℃下孵育4 h后，每个细胞分别摄取了7.2、13.2和27.3 pg的磁铁矿。进一步.在24小时孵育期后未观察到毒性。使用磁性标记的CM（磁铁矿浓度，100 pg/细胞），构建多层CM片。连接蛋白43的免疫荧光染色证明了由Mag-TE构建的CM片层内存在间隙连接。此外，使用细胞外电位标测证实了由Mag-TE构建的CM片内的电连接。这些结果表明，Mag-TE是一种可行的心脏组织工程方法。
Heart tissue engineering requires construction of three-dimensional (3-D) tissues composed of cardiomyocytes (CMs) that are tightly connected to each other. The aim of this study was to construct "scaffold-less" multi-layered 3-D CM sheets using magnetic force-based tissue engineering (Mag-TE) and to evaluate the cell-to-cell functional connections within the CM sheets. Original magnetite cationic liposomes (MCLs) with a positive surface charge (which facilitate adsorption to the target cell surface) were taken up by CMs that were isolated from 2-day-old Wistar rats. When MCLs were added to the medium of CMs at magnetite concentrations of 25, 50, and 100 pg per cell, subsequent measurements showed that 7.2, 13.2, and 27.3 pg of magnetite were taken up per cell, respectively, after 4 h incubation at 37 degrees C. Further. no toxicity was observed after a 24 h incubation period. Using magnetically labeled CMs (magnetite concentration, 100 pg/cell), multi-layered CM sheets were constructed. Immunofluorescent staining of connexin43 demonstrated the presence of gap junctions within the CM sheets that were constructed by Mag-TE. Moreover, electrical connections within the CM sheets constructed by Mag-TE were confirmed using extracellular potential mapping. These results indicate that Mag-TE is a viable methodology for heart tissue engineering.