Construction of heterotypic cell sheets by magnetic force-based 3-D coculture of HepG2 and NIH3T3 cells
Construction of heterotypic cell sheets by magnetic force-based 3-D coculture of HepG2 and NIH3T3 cells
复制标题
DOI：
10.1263/jbb.104.371
复制
发表时间：
2007-11-01
影响因子：
2.8
通讯作者：
Karnihira, Masamichi
中科院分区：
文献类型：
--
作者：
Ito, Akira;Jitsunobu, Hideaki;Karnihira, Masamichi
异型3-D共培养对于模拟组织和器官是必不可少的，因为各种类型细胞之间的细胞-细胞相互作用被认为对于细胞功能的激活是重要的。在这项研究中，施加磁力，以建立一个三维共培养系统的HepG 2和NIH 3 T3细胞作为模型的肝细胞和间充质细胞。磁铁矿阳离子脂质体（MCL）用于标记靶细胞。将用MCL标记的NIH 3 T3细胞接种到超低附着板上，其表面由亲水性和中性电荷的共价结合的水凝胶层组成。当将磁体放置在板下时，细胞在孔的底部积累。经过24小时的孵育期，细胞形成了一个多层的细胞片，其中含有主要的间充质细胞外基质（ECM）成分（纤连蛋白和I型胶原），这表明使用基质NIH 3 T3细胞给予足够的强度细胞片。NIH 3 T3和HepG 2细胞均用MCL标记，并通过两种方法共培养：构建NIH 3 T3细胞片，随后将HepG 2细胞接种到NIH 3 T3细胞片上，然后施加磁力形成分层细胞片;或将NIH 3 T3和HepG 2细胞混合，然后施加磁力形成混合细胞片。成功构建了这些异型多层细胞片，并观察到HepG 2细胞的白蛋白分泌增强。这些结果表明，新的组织工程技术，使用磁铁矿纳米粒子和磁力，我们称之为基于磁力的组织工程（Mag-TE），是一种很有前途的方法来构建由异型共培养细胞组成的多层细胞片。
Heterotypic 3-D coculture is essential to mimic tissues and organs, because cell-cell interaction between various types of cells is believed to be important for the activation of cellular functions. In this study, magnetic force was applied to construct a 3-D coculture system of HepG2 and NIH3T3 cells as a model of hepatocytes and mesenchymal cells. Magnetite cationic liposomes (MCLs) were used to label target cells. NIH3T3 cells labeled with MCLs were seeded onto ultralow-attachment plates, whose surface is composed of a covalently bound hydrogel layer that is hydrophilic and neutrally charged. When a magnet was placed under the plate, cells accumulated on the bottom of the well. After a 24-h incubation period, the cells formed a multilayered cell sheet, which contained the major mesenchymal extracellular matrix (ECM) components (fibronectin and type I collagen), suggesting that the use of stromal NIH3T3 cells gave sufficient strength to cell sheets. Both NIH3T3 and HepG2 cells were labeled with MCLs, and cocultured by two methods: NIH3T3 cell sheets were constructed and HepG2 cells were subsequently seeded onto NIH3T3 cell sheets, and then allowed to form layered cell sheets by applying magnetic force; or NIH3T3 and HepG2 cells were mixed and then allowed to form mixed cell sheets by applying magnetic force. These heterotypic multilayered cell sheets were successfully constructed and an enhanced albumin secretion by HepG2 cells was observed. These results suggest that the new tissue engineering technique using magnetite nanoparticles and magnetic force, to which we refer to as magnetic force-based tissue engineering (Mag-TE), is a promising approach to construct multilayered cell sheets consisting of heterotypic cocultured cells.