Journal
NATURE PROTOCOLS
Volume 8, Issue 10, Pages 1940-1949Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2013.125
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Funding
- US National Science Foundation (NSF) Small Business Innovation Research Award Phase I [0945954]
- NSF IIP Division of Industrial Innovation and Partnerships, and by an award from the State of Texas Emerging Technology [1127551]
- State of Texas Emerging Technology Fund
- Directorate For Engineering
- Div Of Industrial Innovation & Partnersh [1127551, 0945954] Funding Source: National Science Foundation
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Recently, biomedical research has moved toward cell culture in three dimensions to better recapitulate native cellular environments. This protocol describes one method for 3D culture, the magnetic levitation method (MLM), in which cells bind with a magnetic nanoparticle assembly overnight to render them magnetic. When resuspended in medium, an external magnetic field levitates and concentrates cells at the air-liquid interface, where they aggregate to form larger 3D cultures. The resulting cultures are dense, can synthesize extracellular matrix (ECECM) and can be analyzed similarly to the other culture systems using techniques such as immunohistochemical analysis (IHC), western blotting and other biochemical assays. This protocol details the MLM and other associated techniques (cell culture, imaging and IHC) adapted for the MLM. The MLM requires 45 min of working time over 2 d to create 3D cultures that can be cultured in the long term (> 7 d).
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