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In Cell Culture, the Neighbourhood Matters

Photo: A Neighbourhood
Neighbourhood matters – not
only for humans but also for
cells in lab dishes; © panther-
media.net/William Bacon

"Soluble factors are important, but cells normally interact with the extracellular matrix and with neighbouring cells, and these have not been considered in most efforts to refine growth conditions," says Kiessling. "We wanted to know, can we replace the neighbouring cells and extracellular matrix with synthetics?"

Creating a more precise system for growing cells offers both theoretical and practical advantages, Kiessling says. First, it would reduce uncertainty in experiments by simplifying conditions. Second, it would remove the risk of biological contamination like viruses, so the cultured cells could be used in medicine. Third, new surfaces that improve the control over cell growth and development could facilitate the formation of artificial tissues, which are complex assemblies of different cell types.

Kiessling outlines two areas of progress from her lab at UW-Madison. One series of experiments used a lab dish decorated with molecules called peptides to amplify the response of cells to a growth factor called transforming growth factor beta (TGF-beta). TGF-beta can affect healing, cell division and transformation into a more specialized cell or into a tumour cell, so TGF-beta can be helpful or harmful in different situations.

After screening thousands of potential anchors, Kiessling and colleagues found a peptide that would safely hold the cells in place and simultaneously make them extremely sensitive to TGF-beta made by the cells themselves. "TGF-beta receptors have to assemble into a complex before TGF-beta sends its signal to the cell," says Kiessling. "We made surfaces that organized the receptors so they were especially sensitive to this growth factor when the cells were bound to the surface, and so the growth factor affects the cell at incredibly low levels, levels we cannot detect."

The secret, she says, lies in the preparation: The manufactured surface primes the cells to respond to tiny amounts of growth factor. "It's like the surface acts as an amplifier to allow the cells to sense the presence of the growth factor."

When they are grown on surfaces without the peptides, the cells used in the experiment are like skin cells, but when they are on the peptide surface, they detect the growth factor and transform into muscle-like cells. "That shows the power of this approach," says Kiessling. "We have a way to make cells do one thing if they are attached to this surface and another thing if they are not."

A second series of experiments concerned human embryonic stem cells. Their therapeutic potential has remained tantalizingly difficult to reach, partly because they have been grown with substances derived from mice that could contain viruses or other pathogens.

While scientists have refined the liquid portion of the environment needed to grow and transform embryonic stem cells, the solid portion has received less attention. "Human embryonic stem cells need not only a defined medium, but also a defined substrate," Kiessling says. "Historically, the field has relied on mouse embryonic feeder cells and various mixtures of proteins isolated from mice, which contain who knows what in the way of viruses or other infectious particles."

"Our work highlights the fact that we can use a patterned surface itself to instruct the cells, which could be really useful for growing cells on a larger scale and differentiating them under defined conditions," Kiessling says. "Patterned surfaces are found throughout our tissues. Hair, eyes, brain, everywhere we have organized tissues, so if we want to grow different types of cells in ordered arrays to build up a tissue, we need the cells to be organized. We are not close to building a complicated tissue, but the first step is to localize specific types of cells where we want them. We've only scratched the surface of our ability to regulate and instruct cell growth and transformation using elaborately structured surfaces."


MEDICA.de; Source: University of Wisconsin-Madison

 
 

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