Colony, hanging drop, and methylcellulose three dimensional hypoxic growth optimization of renal cell carcinoma cell lines.
Colony, hanging drop, and methylcellulose three dimensional hypoxic growth optimization of renal cell carcinoma cell lines.
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DOI：
10.1007/s10616-016-0063-2
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发表时间：
2017-08
期刊：
影响因子：
2.2
通讯作者：
Czarnecka AM
中科院分区：
文献类型：
--
作者：
Matak D;Brodaczewska KK;Lipiec M;Szymanski Ł;Szczylik C;Czarnecka AM
肾细胞癌（Renal cell carcinoma，RCC）是泌尿系统常见的恶性肿瘤之一，约占人类肿瘤的3%，其发病率呈逐年上升趋势。我们需要新的方法来靶向对当前疗法有抗性并导致复发和治疗失败的肿瘤细胞。在这项研究中，我们专注于低氧张力和三维（3D）细胞培养纳入开发一个新的RCC生长模型。我们使用了3D培养中常见的悬滴和菌落形成方法，以及独特的甲基纤维素（MC）方法。对于实验，我们使用人原代RCC细胞系、转移性RCC细胞系、人肾癌干细胞和人健康上皮细胞。在悬滴试验中，我们验证了各种细胞系在低氧和常氧条件下产生固体聚集体的潜力。利用半软琼脂法，我们还测定了各种细胞系在不同氧条件下产生集落的能力。MC方法与悬滴和集落形成试验中观察到的不同细胞行为表明，这三种试验可用于测试各种细胞特性。然而，MC似乎是3D细胞培养的特别有价值的替代方案，因为其更高的聚集体形成效率和血清不依赖性在癌症生物学的不同领域中是令人感兴趣的。
Renal cell carcinoma (RCC) is the most lethal of the common urologic malignancies, comprising 3% of all human neoplasms, and the incidence of kidney cancer is rising annually. We need new approaches to target tumor cells that are resistant to current therapies and that give rise to recurrence and treatment failure. In this study, we focused on low oxygen tension and three-dimensional (3D) cell culture incorporation to develop a new RCC growth model. We used the hanging drop and colony formation methods, which are common in 3D culture, as well as a unique methylcellulose (MC) method. For the experiments, we used human primary RCC cell lines, metastatic RCC cell lines, human kidney cancer stem cells, and human healthy epithelial cells. In the hanging drop assay, we verified the potential of various cell lines to create solid aggregates in hypoxic and normoxic conditions. With the semi-soft agar method, we also determined the ability of various cell lines to create colonies under different oxygen conditions. Different cell behavior observed in the MC method versus the hanging drop and colony formation assays suggests that these three assays may be useful to test various cell properties. However, MC seems to be a particularly valuable alternative for 3D cell culture, as its higher efficiency of aggregate formation and serum independency are of interest in different areas of cancer biology.
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DOI：
10.4161/cc.8.20.9701
发表时间：
2009-10-15
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Heddleston JM;Li Z;McLendon RE;Hjelmeland AB;Rich JN
通讯作者：
Rich JN
影响因子：
11.2
作者：
Furge, Kyle A.;Chen, Jindong;Teh, Bin Tean
通讯作者：
Teh, Bin Tean
影响因子：
6.4
作者：
Burdett, Emily;Kasper, F. Kurtis;Ludwig, Joseph A.
通讯作者：
Ludwig, Joseph A.
影响因子：
19.7
作者：
Chae, EJ;Kim, JK;Cho, KS
通讯作者：
Cho, KS
影响因子：
6.2
作者：
Dudek, Arkadiusz Z.;Zolnierek, Jakub;Szczylik, Cezary
通讯作者：
Szczylik, Cezary