13.07.2011 -
Contemporary biologists and neuroscientists visualise and study
cells with conventional microscopes with a spatial resolution of 1
micron or slightly less. On the other side, molecular and
structural biologists reason at a molecular level where events and
reactions occur at an Angstrom scale or just above. Therefore,
events occurring at a submicron scale and above the single molecule
dimension represent a new and unexplored perspective for Biology
and Neuroscience.
This scale of analysis was chosen as the focus point for the
work performed by the NanoScale interdisciplinary research
consortium, composed of five major European research centres
(SISSA, CNR-INFM, DTU, NMI and ENS) and three small high-tech
companies (MCS Gmbh, Promoscience Srl, GVT Srl), coordinated by
prof. Vincent Torre from SISSA (Italy).
The project, funded by the European Commission FP7 programme,
has produced an improved map of cell interactions with
nanostructured substrates thanks to new Lab-On-Chip
instruments designed within a partnership between academia and
industry.
These results help the definition of novel cell culture systems,
able to influence specific aspects of cell behaviour, with long
term perspective of application in regenerative therapy of tissues.
The work of the Nanoscale project has been focusing on how a
nanopattern design of cell culture substrates influences
proliferation and differentiation of cells in vitro.
Differentiation is usually achieved by using biochemical
factors. However, biochemical induction does not fully prevent the
presence of some undifferentiated cells that could become
tumorigenic, while the possibility to stimulate and direct neuronal
differentiation by cues encoded in the substrate topography opens
new perspectives for experimental work.
The experiments carried out show that at least one surface
topology nanopatterned in soft elastomer substrates has the
capability to enhance the differentiation of stem cells into
neurons and neural networks: nanopillars. From this point of view,
the scientific results that have been obtained by the project are
not conclusive. The genetic pathways by which the properties of
nanopatterned substrates affect the cell differentiation are still
obscure and need to be analysed more in depth.
On the other side, the technological and methodological
developments achieved by the Nanoscale team show a big potential
for future applications in many research fields.
The partners developed a set of nanotechniques to structure the
surface of cell culture substrates with predefined nanopatterns,
using X-ray lithography and nanoimprinting. MicroElectrode Arrays
(MEAs), which are the most used tool for extracellular in vitro
electrophysiology, have been nanopatterned by nanoimprinting and
integrated into a complete high throughput prototype setup, which
could be commercialised in the future by MultiChannelSystems Gmbh,
one of the NanoScale industrial partners.
In order to find the best pattern configuration for growth
guidance, researchers also tried to apply mix-and-match approaches
of nanoimprinting and conventional photolithography to optimise the
substrates pattern design. T
hey found that while both axons and dendrites follow biochemical
patterns, etched topographic features seem to be more relevant for
axonal guidance. One research team therefore also worked out a
novel patterning technique based on nanoimprint lithography and
reactive ion etch techniques, that can be applied as a general
lithography method alternative to micro-contact print.
The resulting high resolution protein patterns are more suited
for long term cell culture. Moreover, the project team has
developed a novel method for fast 3D imaging of cell interactions
in nanostructures, which allows imaging of nanostructures (e.g.
silicon nanowires) inside the cells.
Finally, a new method for data-driven modelling of cell motility
has been established: it can detect the influence of nanostructures
on cell motility patterns based on statistical analysis of motility
from time lapse movies.
INFO: secretariat@nanoscale-fp7.eu;