Shaping Up

How to find your way around three-dimensional cell culture.

Josh P. Roberts
May 31, 2008
<figcaption>A mammary epithelial cell acinus after eight days of 3D culture. As an acinus develops, the central cells (blue: nuclear staining) occupying the nascent luminal space undergo apoptosis, as depicted by the immunolocalization of activated caspase-3 (green). Credit: Brugge Lab / Harvard Medical School</figcaption>
A mammary epithelial cell acinus after eight days of 3D culture. As an acinus develops, the central cells (blue: nuclear staining) occupying the nascent luminal space undergo apoptosis, as depicted by the immunolocalization of activated caspase-3 (green). Credit: Brugge Lab / Harvard Medical School

Researchers in fields as diverse as neuroscience, angiogenesis, and regenerative medicine are discovering that the surfaces their cultured cells grow on may be as important as the media they're grown in. Cells grown in traditional two-dimensional cultures in flasks and wells have different topology, architecture, and viscoelasticity compared with cells grown in vivo. This affects a host of cell parameters, such as morphology, growth, and polarity. Two-dimensional cultures also neglect the role that the extracellular matrix plays in informing a cell's decisions, as well as in providing a physical space in which to form 3-D structures. "Most phenotypes that we're following now are not revealed in...

Three-dimensional options for cell cultures are out there. The 3-D approach has its challenges, though, beginning with determining which of the many available synthetic and natural matrices best recapitulate your in vivo situation. Some materials may interact with the culture in unpredictable ways. Antibodies used in immunohistochemistry may interact with the matrix. Imaging the culture can be tricky, requiring specialized expertise or equipment to minimize cytotoxic damage. Moreover, although experiments are run in three dimensions, cells are still maintained and passaged in two dimensions.

The Scientist sought out researchers who have perfected their 3-D culturing methods to help you navigate the ins and outs of the technique.

Click on their profiles in the Article Extras box to read about their work.