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首页> 美国卫生研究院文献>Tissue Engineering. Part C Methods >Generation of Homogenous Three-Dimensional Pancreatic Cancer Cell Spheroids Using an Improved Hanging Drop Technique
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Generation of Homogenous Three-Dimensional Pancreatic Cancer Cell Spheroids Using an Improved Hanging Drop Technique

机译：使用改进的悬挂滴技术生成同质三维胰腺癌细胞球体。

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In vitro characterization of tumor cell biology or of potential anticancer drugs is usually performed using tumor cell lines cultured as a monolayer. However, it has been previously shown that three-dimensional (3D) organization of the tumor cells is important to provide insights on tumor biology and transport of therapeutics. Several methods to create 3D tumors in vitro have been proposed, with hanging drop technique being the most simple and, thus, most frequently used. However, in many cell lines this method has failed to form the desired 3D tumor structures. The aim of this study was to design and test an easy-to-use and highly reproducible modification of the hanging drop method for tumor sphere formation by adding methylcellulose polymer. Most pancreatic cancer cells do not form cohesive and manageable spheres when the original hanging drop method is used, thus we investigated these cell lines for our modified hanging drop method. The spheroids produced by this improved technique were analyzed by histology, light microscopy, immunohistochemistry, and scanning electron microscopy. Results show that using the proposed simple method; we were able to produce uniform spheroids for all five of the tested human pancreatic cancer cell lines; Panc-1, BxPC-3, Capan-1, MiaPaCa-2, and AsPC-1. We believe that this method can be used as a reliable and reproducible technique to make 3D cancer spheroids for use in tumor biology research and evaluation of therapeutic responses, and for the development of bio-artificial tissues.
机译：通常使用培养为单层的肿瘤细胞系进行肿瘤细胞生物学或潜在抗癌药物的体外表征。但是，先前已经表明，肿瘤细胞的三维（3D）组织对于提供有关肿瘤生物学和治疗方法的见解很重要。已经提出了几种在体外产生3D肿瘤的方法，其中悬滴技术是最简单的，因此也是最常用的。然而，在许多细胞系中，该方法未能形成所需的3D肿瘤结构。这项研究的目的是通过添加甲基纤维素聚合物来设计和测试一种易于使用且高度可重复的修改悬滴法以形成肿瘤球的方法。当使用原始的悬滴法时，大多数胰腺癌细胞不会形成具有凝聚力且易于管理的球体，因此我们针对改良的悬滴法研究了这些细胞系。通过组织学，光学显微镜，免疫组织化学和扫描电子显微镜分析通过这种改进技术产生的球体。结果表明，该方法简单易行。我们能够为所有五个测试的人类胰腺癌细胞系生产均匀的球体； Panc-1，BxPC-3，Capan-1，MiaPaCa-2和AsPC-1。我们相信，该方法可以用作制造3D癌症球体的可靠且可重现的技术，以用于肿瘤生物学研究和评估治疗反应以及生物人工组织的发展。

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