title: Towards automated production and drug sensitivity testing using scaffold-free spherical tumor microtissues.
abstract: 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.
authors.affiliation: InSphero AG, Zürich, Switzerland.
authors.forename: Maren
authors.initials: M
authors.lastname: Drewitz
authors.forename: Marianne
authors.initials: M
authors.lastname: Helbling
authors.forename: Nicole
authors.initials: N
authors.lastname: Fried
authors.forename: Manuela
authors.initials: M
authors.lastname: Bieri
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Moritz
authors.forename: Jan
authors.initials: J
authors.lastname: Lichtenberg
authors.forename: Jens M
authors.initials: JM
authors.lastname: Kelm
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D055808:Drug Discovery / Q000379:methods*; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D045325:HCT116 Cells; D019073:HT29 Cells; D056945:Hep G2 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D020128:Inhibitory Concentration 50; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology; D018874:Spheroids, Cellular / Q000187:drug effects / Q000473:pathology; D046509:Tissue Culture Techniques / Q000379:methods*