Towards automated production and drug sensitivity testing using scaffold-free spherical tumor microtissues

Biotechnol J. 2011 Dec;6(12):1488-96. doi: 10.1002/biot.201100290. Epub 2011 Nov 21.


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 IC(50) 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.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Drug Discovery / methods*
  • Drug Screening Assays, Antitumor / methods*
  • HCT116 Cells
  • HT29 Cells
  • Hep G2 Cells
  • High-Throughput Screening Assays / methods*
  • Humans
  • Inhibitory Concentration 50
  • Neoplasms / drug therapy
  • Neoplasms / pathology
  • Spheroids, Cellular / drug effects
  • Spheroids, Cellular / pathology
  • Tissue Culture Techniques / methods*