Three-Dimensional In Vitro Hepatic Constructs Formed Using Combinatorial Tapered Stencil for Cluster Culture (TASCL) Device
Three-Dimensional In Vitro Hepatic Constructs Formed Using Combinatorial Tapered Stencil for Cluster Culture (TASCL) Device
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DOI：
10.3727/215517914x685187
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Hayashi, Shuji
中科院分区：
文献类型：
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作者：
Miyamoto, Yoshitaka;Ikeuchi, Masashi;Hayashi, Shuji
从各种细胞中创建人工肝组织的尝试已被报道为肝移植和药物测试的替代方法。在人工肝组织的构建中，细胞来源的选择是最重要的因素。然而，如果不能为各种细胞提供适当的环境（体外/体内），则不可能获得具有所需功能的人工肝组织。因此，我们专注于体外环境，并使用MEMS技术生产肝组织。在本研究中，我们报告了一种组合TASCL装置，以在体外制备3D细胞构建体。TASCL装置被制造成具有10 mm × 10 mm的总体尺寸，具有微孔和每个微孔的顶部孔径（400 μ m × 400 μ m、600 μ m × 600 μ m、800 μ m × 800 μ m）和底部孔径（40 μ m × 40 μ m、80 μ m × 80 μ m、160 μ m × 160 μ m）。TASCL装置可以很容易地用镊子安装在各种培养皿上。使用塑料皿作为组合TASCL装置的底表面，通过增加原代小鼠肝细胞的接种细胞密度来产生均匀尺寸（约φ 100 μ m-φ 200 mm）的3D肝细胞构建体。使用TASCL装置获得的3D肝细胞构建体是活的并且分泌白蛋白。另一方面，使用组合TASCL装置，部分粘附的原代小鼠肝细胞在单个微孔的胶原包被底部上表现出鹅卵石形态。通过改变TASCL装置的底部基底，细胞构建体的培养环境很容易改变为3D环境。本报告中描述的组合TASCL设备可以快速简单地使用。该装置将用于制备肝细胞构建体，以应用于药物筛选和细胞医学。
Attempts to create artificial liver tissue from various cells have been reported as an alternative method for liver transplantation and pharmaceutical testing. In the construction of artificial liver tissue, the selection of the cell source is the most important factor. However, if an appropriate environment (in vitro/in vivo) cannot be provided for various cells, it is not possible to obtain artificial liver tissue with the desired function. Therefore, we focused on the in vitro environment and produced liver tissues using MEMS technology. In the present study, we report a combinatorial TASCL device to prepare 3D cell constructs in vitro. The TASCL device was fabricated with an overall size of 10 mm x 10 mm with microwells and a top aperture (400 mu m x 400 mu m, 600 mu m x 600 mu m, 800 mu m x 800 mu m) and bottom aperture (40 mu m x 40 mu m, 80 mu m x 80 mu m, 160 mu m x 160 mu m) per microwell. The TASCL device can be easily installed on various culture dishes with tweezers. Using plastic dishes as the bottom surface of the combinatorial TASCL device, 3D hepatocyte constructs of uniform sizes (about phi 100 mu m-phi 200 mm) were produced by increasing the seeding cell density of primary mouse hepatocytes. The 3D hepatocyte constructs obtained using the TASCL device were alive and secreted albumin. On the other hand, partially adhered primary mouse hepatocytes exhibited a cobblestone morphology on the collagencoated bottom of the individual microwells using the combinatorial TASCL device. By changing the bottom substrate of the TASCL device, the culture environment of the cell constructs was easily changed to a 3D environment. The combinatorial TASCL device described in this report can be used quickly and simply. This device will be useful for preparing hepatocyte constructs for application in drug screening and cell medicine.