用组合锥形模板进行集群培养(TASCL)装置形成的三维离体肝脏结构。

Y. Miyamoto, M. Ikeuchi, H. Noguchi, T. Yagi, S. Hayashi
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引用次数: 6

摘要

据报道,用各种细胞制造人工肝组织是肝移植和药物试验的一种替代方法。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,我们着眼于体外环境,利用MEMS技术生产肝脏组织。在本研究中,我们报道了一种组合TASCL装置,用于体外制备三维细胞结构。TASCL器件的总尺寸为10 mm × 10 mm,每个微孔有顶部孔径(400µm × 400µm, 600µm × 600µm, 800µm × 800µm)和底部孔径(40µm × 40µm, 80µm × 80µm, 160µm × 160µm)。TASCL装置可以用镊子方便地安装在各种培养皿上。采用塑料培养皿作为组合TASCL装置的底表面,通过增加原代小鼠肝细胞的播种细胞密度,可获得均匀大小(约为100 μm ~ 200 μm)的三维肝细胞构建体。使用TASCL装置获得的三维肝细胞结构是活的,并分泌白蛋白。另一方面,使用组合TASCL装置,部分粘附的小鼠原代肝细胞在单个微孔的胶原包被底部呈现鹅卵石状形态。通过改变TASCL装置的底物,可以很容易地将细胞构建物的培养环境改变为三维环境。本报告所描述的组合式TASCL装置使用简单快捷。该装置可用于制备用于药物筛选和细胞医学的肝细胞构建物。
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Three-Dimensional In Vitro Hepatic Constructs Formed Using Combinatorial Tapered Stencil for Cluster Culture (TASCL) Device.
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 × 10 mm with microwells and a top aperture (400 µm × 400 µm, 600 µm × 600 µm, 800 µm × 800 µm) and bottom aperture (40 µm × 40 µm, 80 µm × 80 µm, 160 µm × 160 µm) 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 ɸ 100 μm-ɸ 200 μm) 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 collagen-coated 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.
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Cell medicine
Cell medicine MEDICINE, RESEARCH & EXPERIMENTAL-
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