Towards automated production and drug sensitivity testing using scaffold-free spherical tumor microtissues
Towards automated production and drug sensitivity testing using scaffold-free spherical tumor microtissues
复制标题
DOI：
10.1002/biot.201100290
复制
发表时间：
2011-12-01
影响因子：
4.7
通讯作者：
Kelm, Jens M.
中科院分区：
文献类型：
--
作者：
Drewitz, Maren;Helbling, Marianne;Kelm, Jens M.
尽管三维（3 - D）培养的相关性多年来已得到认可，并在学术层面得到应用，但其向工业应用的转化却进展缓慢。可靠的高通量技术的发展显然是3 - D模型在工业中应用的先决条件。在这项研究中，在标准的96孔通道机器人平台上评估了96孔悬滴多孔板形式的球形微组织生产和药物测试的稳健性。制备了来自六种不同细胞系的微组织模型，并根据其生长曲线和形态进行了表征，这些模型在至少15天内呈现出高密度的组织样重构和生长。选择结肠癌细胞系HCT116作为模型来评估基于微组织的检测重现性。在三个独立的生产批次中，所生产的微组织的大小差异低于5%。使用两种参考化合物——十字孢碱和苯丁酸氮芥测试了基于微组织的检测的可靠性。在四次独立的药物测试中，计算出的半数抑制浓度（IC50）值与二维多孔板测试进行了对比，显示出相似的一致性。这里介绍的用于以标准96孔形式自动生产各种微组织以进行药效测试的技术，将有助于在药物研发过程的早期阶段应用更多的类器官模型。
Although the relevance of three-dimensional (3-D) culture has been recognized for years and exploited at an academic level, its translation to industrial applications has been slow. The development of reliable high-throughput technologies is clearly a prerequisite for the industrial implementation of 3-D models. In this study the robustness of spherical microtissue production and drug testing in a 96-well hanging-drop multiwell plate format was assessed on a standard 96-well channel robotic platform. Microtissue models derived from six different cell lines were produced and characterized according to their growth profile and morphology displaying high-density tissue-like reformation and growth over at least 15 days. The colon cancer cell line HCT116 was chosen as a model to assess microtissue-based assay reproducibility. Within three individual production batches the size variations of the produced microtissues were below 5%. Reliability of the microtissue-based assay was tested using two reference compounds, staurosporine and chlorambucil. In four independent drug testings the calculated IC50 values were benchmarked against 2-D multiwell testings displaying similar consistency. The technology presented here for the automated production of a variety of microtissues for efficacy testing in a standard 96-well format will aid the implementation of more organotypic models at an early time point in the drug discovery process.