Miyamoto Yoshitaka, Ikeuchi Masashi, Noguchi Hirofumi, Yagi Tohru, Hayashi Shuji
Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine , Showa-ku, Nagoya , Japan.
†Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan; ‡PRESTO, Japan Science and Technology (JST), Saitama, Japan.
Cell Med. 2014 Dec 12;7(2):67-74. doi: 10.3727/215517914X685187. eCollection 2015 Feb 8.
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.
据报道，尝试用各种细胞创建人造肝组织作为肝移植和药物测试的替代方法。在人造肝组织的构建中，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合适的环境（体外/体内），就不可能获得具有所需功能的人造肝组织。因此，我们专注于体外环境，并使用微机电系统（MEMS）技术制造肝组织。在本研究中，我们报告了一种组合式TASCL装置，用于在体外制备三维细胞构建体。TASCL装置的整体尺寸为10毫米×10毫米，带有微孔，每个微孔的顶部孔径为（400微米×400微米、600微米×600微米、800微米×800微米），底部孔径为（40微米×40微米、80微米×80微米、160微米×160微米）。TASCL装置可以用镊子轻松安装在各种培养皿上。以塑料培养皿作为组合式TASCL装置的底面，通过增加原代小鼠肝细胞的接种细胞密度，制备了大小均匀（约ɸ100微米-ɸ200微米）的三维肝细胞构建体。使用TASCL装置获得的三维肝细胞构建体具有活性并分泌白蛋白。另一方面，使用组合式TASCL装置，部分贴壁的原代小鼠肝细胞在各个微孔胶原包被的底部呈现鹅卵石形态。通过改变TASCL装置的底部基质，细胞构建体的培养环境很容易转变为三维环境。本报告中描述的组合式TASCL装置使用起来快速简便。该装置将有助于制备用于药物筛选和细胞医学的肝细胞构建体。
