Award details

Progression of animal cell interactions and intracellular processes triggered on contact in ultrasound standing wave traps

Reference	BB/C515220/1
Principal Investigator / Supervisor	Dr James Ralphs
Co-Investigators / Co-Supervisors	Professor Charles Archer, Professor Wendy Purcell
Institution	Cardiff University
Department	School of Biosciences
Funding type	Research
Value (£)	406,588
Status	Completed
Type	Research Grant
Start date	01/05/2005
End date	30/04/2008
Duration	36 months

Abstract

The PI¿s group has recently shown that two dimensional aggregates of cells in suspension can be monitored microscopically as they form, are held in suspension and then develop into a cell monolayer in an ultrasound trap. The monolayer cell structure provides a high quality environment to exploit the power of fluorescence microscopy in developing cell aggregates. This system will be employed to measure and compare the temporal development of surface receptor adhesiveness, intracellular cytoskeletal development, viability and proliferation of different cell types in aggregates when these are physically and non-intrusively held, for times up to 3 days, away from the constraints of substrata. The aggregate morphology, during and following formation, will be characterised by fractal analysis and other indices. The stability of the aggregates against flow stress and the temporal changes in physical morphology indices as the aggregate undergoes compaction will be correlated with development of the actin cytoskeleton at cell contact regions. This generic 2-D approach will contrast the behaviour of anchorage independent (helatocytes) and anchorage dependent (chinese hamster ovary (CHO) cells and pre-chondrogenic chick embryo) cells. Hepatocytes are of interest in tissue engineering and in multicellular spheroid development as animal-free organotypic test-beds in Toxicology. Pre-chondrogenic cells will be studied from the biophysics and kinetics of initial aggregation through to the molecular biology of the condensation stage. Development of compartmentalisation of cell type within the pre-chondrogenic cell aggregate will be monitored by Voronoi partition analysis. The extent to which proliferation of anchorage dependent CHO cells, a work-horse for large scale cultures in the pharmaceutical industry, can continue in ultrasound aggregates without the medium changes currently required for proliferation of these cells in suspension will be investigated. A second ultrasound standing wave approach will be developed to produce 3-D aggregates of pre-chondrogenic chick embryo cells for differentiation studies. Ultrasound-produced 3-D aggregates of hepatocytes will be tested for their functionality. Their potential as nucleation aggregates for production of functional organotypic spheroids in conventional culture will also be assesses. In the course of addressing the problems outlined above the project will develop techniques that will be immediately applicable in areas such as tumour cell invasion, tissue engineering, systems biology and controlled organisation of cell consortia in vitro.

Summary

unavailable

Committee	Closed Committee - Engineering & Biological Systems (EBS)
Research Topics	Industrial Biotechnology, Regenerative Biology, Technology and Methods Development
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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