WARF: P06285US
Method of Patterning Cells for Cancer Metastasis or Invasion Assays

INVENTORS

•

David Beebe, Vinay Abhyankar

Microfluidic systems provide excellent platforms for studying biological interactions because they enable precise control over cellular positioning and environmental cues in culture. The ability to control the position of cells in a culture allows physiologically relevant cells to be cultured together to more closely recapitulate the in vivo behavior of the target cell population. Many techniques have been developed to effectively position cells; however, it is not currently possible to investigate the effect of sequentially adding new cell populations into spatially defined regions of the culture.

Controlling environmental cues, such as chemical gradients, is useful because gradients play a key role in mediating biological activity in vivo. Cell invasion assays are typically used to determine the migratory characteristics of particular cells, but it is currently not possible to obtain useful information about the characteristic behavior of cells in response to a gradient before or after invasion using available assays. UW-Madison researchers have developed a method for patterning particles, such as cells, on an arbitrary substrate. The method allows for the sequential addition of different particle populations into spatially defined regions of the substrate. It can be used to conduct a real-time quantitative investigation of cell invasion or migration in response to a predictable gradient.

In this method, a suspension that includes the particles is deposited onto the upper end of an access port within a microfluidic device. The access port is one of several that connect the upper surface of the device to a channel within the device. The particles then settle onto and are patterned along the first part of the lower surface of the channel. After a predetermined amount of time, a second suspension that includes particles may be deposited into a second access port so the particles can settle onto and be patterned along a second portion of the channel. To create a gradient, other particles are deposited onto a porous membrane located at the lower end of an access port. The particles then diffuse through the membrane into the channel. 

• Cancer metastis or matrix invasion assays

• Provides an in vitro environment more representative of in vivo conditions 
• Capable of determining the sensitivity of tumor cells to growth factor gradients
• Can sort cells during epithelial-to-mesenchymal transition (EMT)
• Capable of determining what gradient concentration causes the most cells to enter or invade the channel
• Useful to track cell migration after invasion
• Channel may be filled with a fluid or a polmerizable gel.

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