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BioForce Nanosciences to Develop Neural Cell-Based Assays with France’s UPMC, CNRS

BioForce Nanosciences, together with the University Pierre and Marie Curie and the Centre National de Recherche Scientifique in Paris, plan to co-develop neural cell-based assays, a BioForce official said this week.
The assays, which BioForce plans eventually to market to the research community, will be based on BioForces’ Nano eNabler molecular printer, and will be designed to screen candidate treatments for neurodegenerative diseases and neurological trauma, product manager Michael Lynch told CBA News.
“I do not think we have gotten to the specifics of how people will use them, but those are two of the leading examples,” said Lynch. “They will be used to screen compounds and dissect out the mechanisms of those processes.”
As part of the alliance, UPMC and the CNRS will study the structure, function, and microenvironments of neural cells, and BioForce will help the scientists use its eNabler instrument. The company will also be responsible for commercializing any assays or technologies that are developed, said Lynch.
These resulting assays would be an improvement on existing tools that screen candidate treatments for neurodegenerative diseases and neurological trauma, in that they more closely mimic an in vivo environment, the company said.
Financial details of the agreement were not disclosed.
BioForce will work with researchers in the laboratory of Fatiha Nothias at the UPMC and CNRS. The company has been working with Nothias’ group for the last two years, and Nothias and her colleagues were among the first to access the eNabler technology, according to Lynch.
“The eNabler was placed in my lab for technical evaluation [in the first half of 2006],” said Nothias, a principal investigator at UPMC and director of research at CNRS. She presented data at the Neuroscience 2008 meeting, held this week in Washington, DC, describing the response of dorsal root ganglia neuron growth cones on micropatterned ECM substrates.
Nothias declined to discuss the data in detail, citing pending publication, except to say that her team “definitely demonstrated that through [the use of the Nano eNabler], we can bring new insights on cell response, and impact on the growth cone.”
She added that the Nano eNabler is very flexible compared to lithography or other techniques for microcompact printing that use a stamp. The eNabler “allows you to load very little of the molecule of interest on the micropattern, which is not easy to do with other techniques,” said Nothias.
The other advantage is that scientists can create “as many [multicomponent patterns] as you want, which is really great. This was also unavailable with other techniques,” she said.
Lynch said the Nano eNabler is “a microfluidics-assisted contact printer.” The device uses a microfabricated cantilever that has integrated microfluidics to deliver liquids from a reservoir down a channel.

“We definitely demonstrated that through [the use of the Nano eNabler], we can bring new insights on cell response, and impact on the growth cone.”

When the tip of this cantilever touches the surface of the micropatterned substrate, a droplet of the compound of interest is dispensed, and researchers can use the instrument to control the timing of how long the cantilever tip touches the surface to dispense liquid and how much pressure is applied by the tip to the surface.                 
The instrument’s software allows investigators to design their own substrate pattern “on the fly,” said Lynch, so it does not require, as in the case of PDMS microcontact printing, someone to design a photomask and fabricate it in a clean room, and perform all the stamping themselves.
The eNabler allows researchers to design all the patterns themselves, print them, and, based on their results, “go back to the software, punch a few buttons on the screen, and come up with a new pattern based on [the] data,” he said. “It allows for fast iterations of your work.”
The agreement between BioForce and UPMC is fairly open-ended, said Lynch. “There were no terms defined in it; we will see how things progress. We have had a good relationship with [Nothias’ lab] over the last two years, and I do not see any indication that that would change.”
The next step in the project would be to develop specific protocols that can be used on patterned surfaces created on the eNabler, said Lynch, adding that “beyond that it would be the development of the specific surfaces for these neural assays.”
This should not be a long process, he said. “As soon as we incorporate the protocols, determining the patterns should be a fairly simple exercise.”
Lynch declined to assign a specific dollar value to the neuroscience research market, except to describe the market as “extremely large.”
‘Do Their Own Thing’
It is widely agreed in biology that cells in vivo encounter complex microenvironments that are very difficult to replicate in vitro. But according to Lynch, BioForce’s Nano eNabler molecular printer can “dissect out all the components of that [microenvironment] and the effects that these components have” in vitro.
He added that typical cell biology applications use the Nano eNabler to create patterned surfaces that simulate a specific cellular microenvironment.
“This can include patterning extracellular matrix proteins, signaling molecules, peptides, and antibodies in spots and lines ranging from sizes smaller than a single cell to much larger features suitable for multiple cells,” Lynch said.
Also, cells cultured on those surfaces can be used to study cell adhesion, cell motility/migration, stem-cell differentiation, neurite outgrowth, and axon guidance.
The result would enable researchers to observe cells’ responses to the cues from their patterned microenvironment, or they may expose cells to different soluble factors once they have been immobilized.
BioForce has been interested in creating ordered arrays comprising immobilized single cells for high-content screening, said Lynch. Conventional high-content assay data analysis can become difficult as cells overlap and interact with one another, and subpopulations with an interesting phenotype can also get lost or hidden in the noise of the bulk population. 
However, by imaging and characterizing a large population of individual cells adhering at known locations, it becomes much easier to analyze and identify those interesting phenotypes within a heterogeneous population, said Lynch.
He mentioned that BioForce is in the process of developing microtiter plate-based products that can immobilize the cell arrays for high-content screening applications. 
Although researchers could use the Nano eNabler to develop custom patterned surfaces “for those who want to do their own thing,” Lynch said, there would also be kits with standardized patterned surfaces, “something that would be a little more accessible to the average lab.”
In terms of the current agreement, “We would like to start with the work that they have done in Paris to develop these standardized patterned surfaces, and methods for using those patterned surfaces for different types of axon guidance assays, and then turn those into consumable products and kits,” said Lynch.

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