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Project Page – Giuseppina Simone –
Carbo-Beads for 3D cell microfluidic assays
Created 10 January 2009
ACTION PLAN
PROBLEMS IDENTIFIED (PI) / OUTSIDE SKILL REQUIRED (OSR) / RESOLVED (R)
A) Background
B) Specific Aims
C) General Approach
D) Design and Fabrication of chip
E) Future Directions
Background: Cancer is a major cause of mortality, due to the inability to identify and selectively target metastatic tumors. Many tumours of epithelial origin secrete abnormal glycoproteins which may be useful as tumour markers for diagnosis or screening. Although these are commonly detected using monoclonal antibodies these onco-developmental antigens differ from normally expressed glycoproteins mainly in their carbohydrate side chain structure rather than in their protein core. Still, In post-genomic era the biological processes can be understood by studying the roles that glycans play.
Carbohydrates compose a large group of biomolecules with diverse structures and are found largely in the form of glycoconjugates inside or on the surface of cells. For probing glycan–protein interactions, carbohydrate microarrays, which are composed of diverse glycans densely and orderly attached to a solid surface have been realized from…nomi degli autori in the 2002.
Still, Cellular phenotypes largely depend on 3D cellular microenvironments, and the availability of various micro- and nanopatterning techniques offers new tools for the observation and regulation of cellular phenotype changes. It would be an important advance to develop a straightforwardmethod of creating nanofeatured arrays and to use these arrays in studying the consequences of the use of nanoenvironments for cell assays on a single chip.
A) SPECIFIC AIMS
AIM 1 - A)
B) GENERAL APPROACH
Aim 1- Bulk cell-carbobeads
Cells were incubated in presence of carbobeads to define the avidity.
Aim 2 – Manufacture of microfluidic device
Design and manufacture of microfluidic chip to isolate tumoral cells
Potential Problem.
Carbohydrate-carbohydrate Interaction is weak and do not allow to isolate cells by a population
Explanation.
In 1989 Hakomori roposed that the first phase of cell adhesion initiates through a carbohydrate-carbohydrate interaction between these multivalent GSL-domains. This initial step is followed by protein-protein interactions between adhesion receptors and protein-carbohydrate interactions between lectins and glycoconjugates. Embryogenesis, metastasis and other cellular proliferation processes are according to Hakomori, mediated by carbohydrate-carbohydrate (carb-carb) interactions.The experiment success will depend on the interval of time the carbo-carbo bind is the dominant phenomenon, meaning that microfluidics can become an essential tool.
Tests to perform. Improve the bead-cell affinity with a lectin-buffer.
FUTURE DIRECTION.
1. To study the carbohydrate microenvironment effect on the cells behaviour in particula in presence of diseases
2. To isolate tumoral cells from whole blood
3. To build polysaccharide chains in situ, inside the chip.