Protocol

Tissue Engineering

Volume 140 of the series Methods in Molecular Medicine™ pp 141-151

Generation of Multicellular Tumor Spheroids by the Hanging-Drop Method

  • Nicholas E. TimminsAffiliated withDepartment of Surgery and Research, University Hospital Basel
  • , Lars K. NielsenAffiliated withDepartment of Chemical Engineering, The University of Queensland

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Summary

Owing to their in vivo-like characteristics, three-dimensional (3 D) multicellular tumor spheroid (MCTS) cultures are gaining increasing popularity as an in vitro model of tumors. A straightforward and simple approach to the cultivation of these MCTS is the hanging-drop method. Cells are suspended in droplets of medium, where they develop into coherent 3 D aggregates and are readily accessed for analysis. In addition to being simple, the method eliminates surface interactions with an underlying substratum (e.g., polystyrene plastic or agarose), requires only a low number of starting cells, and is highly reproducible. This method has also been applied to the co-cultivation of mixed cell populations, including the co-cultivation of endothelial cells and tumor cells as a model of early tumor angiogenesis.

Key Words

MCTS Multicellular tumor spheroid Tumor Angiogenesis Cancer 3 D cell culture