Comparative Analysis of 3D Bladder Tumor Spheroids Obtained by Forced Floating and Hanging Drop Methods for Drug Screening.
Comparative Analysis of 3D Bladder Tumor Spheroids Obtained by Forced Floating and Hanging Drop Methods for Drug Screening.
复制标题
通过强制浮动和悬挂掉落方法获得药物筛查获得的3D膀胱肿瘤球体的比较分析。
DOI：
10.3389/fphys.2017.00605
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发表时间：
2017
影响因子：
4
通讯作者：
Swiech K
中科院分区：
文献类型：
--
作者：
Amaral RLF;Miranda M;Marcato PD;Swiech K
简介：使用三维（3D）细胞培养物的基于细胞的测定可以更准确地反映化合物的抗肿瘤活性，因为这些模型更好地再现了肿瘤微环境。研究方法：在这里，我们报告了一个比较分析的细胞行为的两个最广泛采用的方法，三维球体培养，强制浮动（超低附着，贴壁，板），和悬滴（HD）的方法，使用RT 4人膀胱癌细胞系作为模型。首先使用4种不同的细胞接种浓度（0.5、1.25、2.5和3.75 × 104个细胞/mL）表征生成的球状体的形态参数和生长/代谢，然后进行耐药性评价。结果如下：无论使用的细胞接种浓度如何，这两种方法都在约48小时的球化时间内产生具有光滑表面和圆形形状的球状体。与二维（2D）培养物相比，在3D培养物中观察到细胞生长和代谢减少。使用0.5和1.25 × 104个细胞/mL的初始培养物（对于荧光法）和2.5和3.75 × 104个细胞/mL的初始培养物（对于HD法）获得球状体直径的最佳范围（300-500 μm）。与2D培养物（IC 50范围为0.39至0.43）相比，在3D条件下培养的RT 4细胞也表现出对多柔比星的更高抗性（IC 50分别为1.00和0.83 μg/mL）。结论：比较结果，我们得出结论，使用平板的强制漂浮方法被认为更适合和直接地产生用于药物筛选/细胞毒性测定的RT 4球状体。这里提出的结果也有助于提高广泛应用所需的3D培养物的标准化。
Introduction: Cell-based assays using three-dimensional (3D) cell cultures may reflect the antitumor activity of compounds more accurately, since these models reproduce the tumor microenvironment better. Methods: Here, we report a comparative analysis of cell behavior in the two most widely employed methods for 3D spheroid culture, forced floating (Ultra-low Attachment, ULA, plates), and hanging drop (HD) methods, using the RT4 human bladder cancer cell line as a model. The morphology parameters and growth/metabolism of the spheroids generated were first characterized, using four different cell-seeding concentrations (0.5, 1.25, 2.5, and 3.75 × 104 cells/mL), and then, subjected to drug resistance evaluation. Results: Both methods generated spheroids with a smooth surface and round shape in a spheroidization time of about 48 h, regardless of the cell-seeding concentration used. Reduced cell growth and metabolism was observed in 3D cultures compared to two-dimensional (2D) cultures. The optimal range of spheroid diameter (300–500 μm) was obtained using cultures initiated with 0.5 and 1.25 × 104 cells/mL for the ULA method and 2.5 and 3.75 × 104 cells/mL for the HD method. RT4 cells cultured under 3D conditions also exhibited a higher resistance to doxorubicin (IC50 of 1.00 and 0.83 μg/mL for the ULA and HD methods, respectively) compared to 2D cultures (IC50 ranging from 0.39 to 0.43). Conclusions: Comparing the results, we concluded that the forced floating method using ULA plates was considered more suitable and straightforward to generate RT4 spheroids for drug screening/cytotoxicity assays. The results presented here also contribute to the improvement in the standardization of the 3D cultures required for widespread application.
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影响因子：
--
作者：
Daoud G;Monzer A;Bahmad H;Chamaa F;Hamdar L;Mouhieddine TH;Shayya S;Eid A;Kobeissy F;Liu YN;Abou-Kheir W
通讯作者：
Abou-Kheir W
影响因子：
3.7
作者：
Ivanov DP;Parker TL;Walker DA;Alexander C;Ashford MB;Gellert PR;Garnett MC
通讯作者：
Garnett MC
影响因子：
3.7
作者：
Hongisto V;Jernström S;Fey V;Mpindi JP;Kleivi Sahlberg K;Kallioniemi O;Perälä M
通讯作者：
Perälä M
影响因子：
3.7
作者：
Huang H;Parlier R;Shen JK;Lutty GA;Vinores SA
通讯作者：
Vinores SA
影响因子：
3.7
作者：
Conde, Vanessa R.;Oliveira, Pedro E.;Silva, Branca M.
通讯作者：
Silva, Branca M.