Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types
Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types
复制标题
DOI：
10.1002/bit.10655
复制
发表时间：
2003-07-20
影响因子：
3.8
通讯作者：
Nielsen, LK
中科院分区：
文献类型：
--
作者：
Kelm, JM;Timmins, NE;Nielsen, LK
多细胞肿瘤球体（MCTS）被用作正常和实体瘤组织的器官型模型。用于产生MCTS的传统技术，例如在非粘附表面上、在悬浮液中或在支架上生长，具有许多缺点，包括需要手动选择以实现均质群体和使用非生理基质化合物。在这项研究中，我们描述了一种温和的方法，产生MCTS，其中个别球状体形式的悬滴悬浮在微量滴定板。该方法已成功应用于广泛的细胞系，并显示出接近100%的效率（即，每滴一个球状体）。使用肝癌细胞系HepG 2，悬滴法产生了具有窄粒度分布的圆形MCTS（变异系数[CV]为10%至15%，而在非粘附表面上生长的变异系数[CV]为40%至60%）。HepG 2和乳腺癌细胞系MCF-7的结构分析，组成的球体，揭示了高度组织化的，三维的，组织样结构与广泛的细胞外基质。悬滴法代表了MCTS生产的一种有吸引力的替代方法，因为它温和，可应用于多种细胞系，并且可产生均匀大小的球状体，而无需筛分或手动选择。该方法可应用于生理学和代谢、肿瘤生物学、毒理学、细胞组织和生物人工组织的发展的基础研究。(C)2003 Wiley Periodicals，Inc.
Multicellular tumor spheroids (MCTS) are used as organotypic models of normal and solid tumor tissue. Traditional techniques for generating MCTS, such as growth on nonadherent surfaces, in suspension, or on scaffolds, have a number of drawbacks, including the need for manual selection to achieve a homogeneous population and the use of nonphysiological matrix compounds. In this study we describe a mild method for the generation of MCTS, in which individual spheroids form in hanging drops suspended from a microtiter plate. The method has been successfully applied to a broad range of cell lines and shows nearly 100% efficiency (i.e., one spheroid per drop). Using the hepatoma cell line, HepG2, the hanging drop method generated well-rounded MCTS with a narrow size distribution (coefficient of variation [CV] 10% to 15%, compared with 40% to 60% for growth on nonadherent surfaces). Structural analysis of HepG2 and a mammary gland adenocarcinoma cell line, MCF-7, composed spheroids, revealed highly organized, three-dimensional, tissue-like structures with an extensive extracellular matrix. The hanging drop method represents an attractive alternative for MCTS production, because it is mild, can be applied to a wide variety of cell lines, and can produce spheroids of a homogeneous size without the need for sieving or manual selection. The method has applications for basic studies of physiology and metabolism, tumor biology, toxicology, cellular organization, and the development of bioartificial tissue. (C) 2003 Wiley Periodicals, Inc.