A New In Vitro Co-Culture Model Using Magnetic Force-Based Nanotechnology.
A New In Vitro Co-Culture Model Using Magnetic Force-Based Nanotechnology.
复制标题
使用基于磁力的纳米技术的新体外共培养模型。
DOI：
10.1002/jcp.25342
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发表时间：
2016
影响因子：
5.6
通讯作者：
Lee JK.
中科院分区：
文献类型：
--
作者：
Takanari H;Miwa K;Fu X;Nakai J;Ito A;Ino K;Honda H;Tonomura W;Konishi S;Opthof T;van der Heyden MA;Kodama I;Lee JK.
成肌细胞（SkMB）移植已作为严重心力衰竭的治疗策略进行。然而，移植后的致瘤性仍然没有解决。我们开发了一种成肌细胞移植的体外模型，该模型采用SkMB和心肌细胞的“模式化”或“随机混合”共培养，从而能够进行后续的电生理学和致心律失常评价。用磁铁矿纳米颗粒磁性标记SkMB，并在多电极阵列上与新生大鼠心室肌细胞（NRVM）共培养。SkMB由阵列下方的磁铁形成图案。使用基因编码的Ca 2+指示剂G-CaMP 2通过Ca 2+成像评价激发同步性。在NRVM的单一培养物中（对照），传导组织良好。在NRVM和SkMB（随机组）的随机混合共培养中，存在来自多个来源的不均匀传导。在“模式化”共培养中，将整体SKMB层插入NRVM层，尽管传导被SkMB区域扭曲，但兴奋均匀传播。随机组的4 mm距离传导时间（CT）（197 ± 126 ms）显著长于对照组（17 ± 3 ms）。在模式组中，通过NRVM-区域的CT没有变化（25 ± 3 ms），尽管通过SkMB-区域的CT显著更长（132 ± 77 ms）。在随机组中，自发兴奋之间的间隔在心跳与心跳之间变化，而在对照组和模式组中记录了规则的心跳。在模式组中观察到NRVMs的同步Ca 2+瞬变，而在随机组中观察到的是异步的。SkMB的图案化排列与磁性纳米颗粒是可行的。使用新的体外模型模拟细胞移植，它可能成为可能的预测由于异源细胞移植的致瘤性。J.细胞。231：2249-2256，2016。© 2016 Wiley Periodicals，Inc.
Skeletal myoblast (SkMB) transplantation has been conducted as a therapeutic strategy for severe heart failure. However, arrhythmogenicity following transplantation remains unsolved. We developed an in vitro model of myoblast transplantation with “patterned” or “randomly‐mixed” co‐culture of SkMBs and cardiomyocytes enabling subsequent electrophysiological, and arrhythmogenic evaluation. SkMBs were magnetically labeled with magnetite nanoparticles and co‐cultured with neonatal rat ventricular myocytes (NRVMs) on multi‐electrode arrays. SkMBs were patterned by a magnet beneath the arrays. Excitation synchronicity was evaluated by Ca2+imaging using a gene‐encoded Ca2+indicator, G‐CaMP2. In the monoculture of NRVMs (control), conduction was well‐organized. In the randomly‐mixed co‐culture of NRVMs and SkMBs (random group), there was inhomogeneous conduction from multiple origins. In the “patterned” co‐culture where an en bloc SKMB‐layer was inserted into the NRVM‐layer, excitation homogenously propagated although conduction was distorted by the SkMB‐area. The 4‐mm distance conduction time (CT) in the random group was significantly longer (197 ± 126 ms) than in control (17 ± 3 ms). In the patterned group, CT through NRVM‐area did not change (25 ± 3 ms), although CT through the SkMB‐area was significantly longer (132 ± 77 ms). The intervals between spontaneous excitation varied beat‐to‐beat in the random group, while regular beating was recorded in the control and patterned groups. Synchronized Ca2+transients of NRVMs were observed in the patterned group, whereas those in the random group were asynchronous. Patterned alignment of SkMBs is feasible with magnetic nanoparticles. Using the novel in vitro model mimicking cell transplantation, it may become possible to predict arrhythmogenicity due to heterogenous cell transplantation. J. Cell. Physiol. 231: 2249–2256, 2016. © 2016 Wiley Periodicals, Inc.