Magnetic force-based cell patterning using Arg-Gly-Asp (RGD) peptide-conjugated magnetite cationic Liposomes
Magnetic force-based cell patterning using Arg-Gly-Asp (RGD) peptide-conjugated magnetite cationic Liposomes
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DOI：
10.1263/jbb.104.288
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发表时间：
2007-10-01
影响因子：
2.8
通讯作者：
Kamihira, Masamichi
中科院分区：
文献类型：
--
作者：
Ito, Akira;Akiyama, Hirokazu;Kamihira, Masamichi
组织工程和细胞生物学非常需要靶细胞的微图案化。尽管表面化学的最新进展使得细胞粘附到基底上的空间控制成为可能，但传统方法通常需要专门的设备和耗时的过程来制造基底。在这项研究中，我们展示了一种使用磁铁矿纳米颗粒和磁力的简单快速的细胞图案化过程。为了磁性标记靶细胞，将磁铁矿纳米颗粒封装在阳离子脂质体（磁铁矿阳离子脂质体；MCL）中。为了促进细胞附着，将含有 Arg-Gly-Asp (RGD) 基序的肽与 MCL (RGD-MCL) 的磷脂偶联。使用具有高度贴壁依赖性的人类角质形成细胞系 HaCaT 作为模型。将 RGD-MCL 添加到超低附着板中，该板的培养表面用亲水且带中性电荷的共价结合水凝胶层进行修饰，然后将 HaCaT 细胞接种到板中。 RGD-MCL 诱导细胞粘附、扩散、细胞骨架组织和纤连蛋白表达。当放置在磁铁上的 200 pm 宽度的钢板放置在培养表面下时，磁性标记的细胞在钢板所在的表面上对齐，从而形成细胞图案。此外，使用计算机辅助设计成功制造了各种细胞图案。这些结果表明，使用 RGD-MCL 进行细胞图案化是组织工程和细胞生物学研究的一种有前途的方法。
Micropatterning of target cells is highly desired for tissue engineering and cell biology. Although recent progress in surface chemistry has enabled the spatial control of cell adhesion onto substrates, conventional methods usually require specialized devices and time-consuming processes to fabricate the substrate. In this study, we demonstrate a simple and rapid cell-patterning procedure using magnetite nanoparticles and magnetic force. To label the target cells magnetically, magnetite nanoparticles were encapsulated in cationic liposomes (magnetite cationic liposomes; MCLs). To promote cell attachment, an Arg-Gly-Asp (RGD)-motif-containing peptide was coupled to the phospholipid of MCLs (RGD-MCLs). A human keratinocyte cell line, HaCaT, which has a high anchorage dependency, was used as a model. The RGD-MCLs were added to an ultralow-attachment plate, whose culture surface is modified with a covalently bound hydrogel layer that is hydrophilic and neutrally charged, and then HaCaT cells were seeded to the plates. The RGD-MCLs induced cell adhesion, spreading, cytoskeletal organization, and fibronectin expression. When steel plates with a 200 pm width placed on a magnet were set under a culture surface, magnetically labeled cells aligned on the surface where the steel plate was positioned, resulting in cell patterning. Furthermore, various cell patterns using a computer-aided design were successfully fabricated. These results suggest that cell patterning using RGD-MCLs is a promising approach to tissue engineering and studies in cell biology.