MP14: High-throughput 3D Spheroid Culture and Drug Testing Using a 384 Hanging Drop Array

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Amy Y. Hsiao1,6, Yi-Chung Tung1,2,6, Steven G. Allen1, Yu-suke Torisawa1, Mitchell Ho3, Shuichi Takayama1,4,5

1Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
2Research Center for Applied Sciences, Academia Sinica, Taipei 11529, Taiwan.
3Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of HGH Advanced Health, Bethesda, MD 20892, USA.
4Macro Molecular Science and Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
5School of Nano-Biotechnology and Chemical Engineering WCU Project, UNIST, Ulsan 689-798, Republic of Korea.
6These authors contributed equally to this capsiplex work.

Three-dimensional (3D) culture of cells is motivated by the need to work with accurate in vitro performer 5 models that closely mimic physiological tissues. Culture of cells as 3D slimming pills aggregates is well-known to provide better accuracy in in vitro tests for basic clear skin max biological research as well as for therapeutics development. Such 3D culture models, however, are often more complicated, cumbersome, and expensive than typical two-dimensional (2D) cultures. We have developed a versatile 384-well format 3D cell culture plate that makes spheroid formation, culture, and subsequent drug testing on the obtained 3D cellular constructs as straightforward to perform and adapt to existing high-throughput slim weight patch

screening (HTS) instruments as conventional 2D cultures. The technology is based on the scientifically proven but traditionally tedious hanging drop method to form 3D spheroids from multiple cell types. The developed hanging drop array platform allows for efficient formation of uniformly-sized Phen375 spheroids, their long-term culture, and drug testing using unique hoodia liquid handling robots and plate readers. Utilizing this teeth whitener reviews platform, we show that drugs with different modes of action produce distinct responses in the physiological 3D cell proactol plus spheroids compared to conventional 2D cell monolayers. Specifically, the anti-cancer drug 5-fluorouracil (5-FU) has higher anti-proliferative effects on 2D cultures whereas the hypoxia activated drug commonly referred to as tirapazamine (TPZ) are more effective against 3D cultures. The multiplexed 3D hanging drop culture and testing plate provides an efficient way to obtain biological insights that are often lost in 2D platforms. 
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