Iijima Yuta, Uenaka Norino, Morimoto Mayu, Sato Daiki, Hirose Satomi, Sakitani Naoyoshi, Shinohara Masahiro, Funamoto Kenichi, Hayase Gen, Yoshino Daisuke
Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-Cho, Koganei, Tokyo 184-8588 Japan.
Faculty of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-Cho, Koganei, Tokyo 184-8588 Japan.
In Vitro Model. 2024 Feb 13;3(1):19-32. doi: 10.1007/s44164-024-00066-3. eCollection 2024 Feb.
Engineered three-dimensional (3D) tissue culture platforms are useful for reproducing and elucidating complex in vivo biological phenomena. Spheroids, 3D aggregates of living cells, are produced based on physicochemical or microfabrication technologies and are commonly used even in cancer pathology research. However, conventional methods have difficulties in constructing 3D structures depending on the cell types, and require specialized techniques/lab know-how to reproducibly control the spheroid size and shape. To overcome these issues, we have developed a fabrication method, which enables anyone to make and mature cancer spheroids using a superhydrophobic microwell made of the monolithic porous materials. Here, we characterize the biological behaviors of the breast cancer spheroids fabricated by our method under normoxic and hypoxic conditions. We found that the fabricated spheroid contracted to a certain size via activation of the actomyosin system. Cell proliferation induced a hypoxic state inside the spheroid (elevated expression of the hypoxia-inducible factor HIF-1α), followed by the formation of a necrotic core and cell escape from the spheroid. In addition, we observed a decrease in cancer spheroid contractility and cell escape from spheroids under hypoxic conditions compared to normoxic conditions, which were related to oxygen concentration-dependent cell motility. The fabricated spheroids perform as 3D tumor tissues in a highly reproducible manner and within a short culture period. Our findings indicate that this fabrication method has a wide range of applications in cancer research, such as elucidating the mechanisms of tumor invasion and metastasis and screening anticancer drugs, as with previous methods.
The online version contains supplementary material available at 10.1007/s44164-024-00066-3.
工程化三维（3D）组织培养平台有助于重现和阐明复杂的体内生物学现象。球体，即活细胞的3D聚集体，是基于物理化学或微制造技术产生的，甚至在癌症病理学研究中也常用。然而，传统方法在根据细胞类型构建3D结构方面存在困难，并且需要专门技术/实验室知识来可重复地控制球体大小和形状。为克服这些问题，我们开发了一种制造方法，任何人都可以使用由整体多孔材料制成的超疏水微孔来制造并使癌症球体成熟。在此，我们表征了通过我们的方法在常氧和低氧条件下制造的乳腺癌球体的生物学行为。我们发现，制造的球体通过肌动球蛋白系统的激活收缩到一定大小。细胞增殖在球体内诱导低氧状态（缺氧诱导因子HIF-1α表达升高），随后形成坏死核心并使细胞从球体中逸出。此外，我们观察到与常氧条件相比，低氧条件下癌症球体的收缩性降低且细胞从球体中逸出，这与氧浓度依赖性细胞运动性有关。制造的球体在短培养期内以高度可重复的方式表现为3D肿瘤组织。我们的研究结果表明，这种制造方法在癌症研究中有广泛应用，例如阐明肿瘤侵袭和转移机制以及筛选抗癌药物，与先前方法一样。
在线版本包含可在10.1007/s44164-024-00066-3获取的补充材料。
