Novel 3D Liquid Cell Culture Method for Anchorage-independent Cell Growth, Cell Imaging and Automated Drug Screening.
Novel 3D Liquid Cell Culture Method for Anchorage-independent Cell Growth, Cell Imaging and Automated Drug Screening.
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DOI：
10.1038/s41598-018-21950-5
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发表时间：
2018-02-26
影响因子：
4.6
通讯作者：
Nishino T
中科院分区：
文献类型：
--
作者：
Abe-Fukasawa N;Otsuka K;Aihara A;Itasaki N;Nishino T
在三维（3D）培养物中生长的细胞比在对细胞施加机械约束的2D培养物中更可能具有天然细胞-细胞和细胞-基质相互作用。然而，大多数3D培养物利用凝胶基质，其虽然用作支架，但由于其固体和不透明的性质以及细胞暴露于外源信号的不一致性而限制了应用。在没有凝胶基质的3D培养中，细胞倾向于彼此粘附并形成中心有坏死区的团块，使其不适合分析。在这里，我们报告称，添加低分子量琼脂命名为LA 717培养基允许细胞生长为分散的克隆球体在3D。LA 717保持细胞分散并沉降到培养基底部，同时保持培养基澄清，几乎没有额外的粘度，使其适合显微镜观察。重要的是，在含LA 717的培养基中形成的癌球状体对抗癌药物如曲美替尼和MK-2206显示出更高的敏感性，这些药物在2D中不那么有效。由于球状体的小且一致的尺寸，细胞活力和药物毒性在自动成像分析中容易检测。这些结果表明，LA 717提供了一种新的三维培养系统，具有很大的体内反射和实用性。
Cells grown in three-dimensional (3D) cultures are more likely to have native cell-cell and cell-matrix interactions than in 2D cultures that impose mechanical constraints to cells. However, most 3D cultures utilise gel matrix which, while serving as a scaffold, limits application due to its solid and opaque nature and inconsistency in cell exposure to exogenous signals. In 3D culture without gel matrix, cells tend to adhere to each other and form clumps with necrotic zone at the centre, making them unsuitable for analyses. Here we report that addition of low-molecular-weight agar named LA717 to culture media allows cells to grow as dispersed clonal spheroids in 3D. LA717 maintains cells dispersed and settled to the bottom of the medium while keeping the medium clear with little additional viscosity, making it suitable for microscopic observation. Importantly, cancer spheroids formed in LA717-containing medium show higher sensitivity to anti-cancer drugs such as Trametinib and MK-2206 that are not as effective in 2D. Because of the small and consistent size of spheroids, cell viability and drug toxicity are readily detectable in automated imaging analysis. These results demonstrate that LA717 offers a novel 3D culture system with great in vivo reflection and practicality.
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影响因子：
4
作者：
Mizushima, Hiroto;Wang, Xiaobiao;Mekada, Eisuke
通讯作者：
Mekada, Eisuke
影响因子：
4.6
作者：
Rhim, Ji Heon;Luo, Xiangjian;Wong, Stephen T. C.
通讯作者：
Wong, Stephen T. C.
影响因子：
8.8
作者：
LIEBMANN, JE;COOK, JA;LIPSCHULTZ, C;TEAGUE, D;FISHER, J;MITCHELL, JB
通讯作者：
MITCHELL, JB
影响因子：
--
作者：
Bokobza SM;Jiang Y;Weber AM;Devery AM;Ryan AJ
通讯作者：
Ryan AJ
影响因子：
1.2
作者：
Borowicz, Stanley;Van Scoyk, Michelle;Winn, Robert A.
通讯作者：
Winn, Robert A.