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3D CO-CULTURE OF PODOCYTES AND ENDOTHELIAL CELLS
Summary
Cells live in a three dimensional (3D) microenviroment which
deeply influences their function.
The present invention relates to a new 3D co-culture method of
podocytes and endothelial cells and a relative co-culture system
and the use of this co-culture system as an in vitro study model
of pathologies affecting the kidneys, and in particular the renal
glomerular filtration barrier.
Technology description.
Although classical culture methods are essential to define basic
properties of single cell types, cells live in a three-dimensional
microenvironment and particularly, podocytes in the glomerulus
make adhesions to the external side of the glomerular basement
membrane, where endothelial cells line the internal side. Podocytes,
endothelial cells and the intervening glomerular basement membrane
form together the glomerular filtration barrier. Though the need of coculturing podocytes and endothelial cells is widely recognized in the
nephrology research community, this is the first successful 3D
system for culturing them in vitro.
Competitive advantages.
With our method, podocytes and endothelial cells are grown on a
isoporous membrane, covering the external and internal side
respectively. Thus, cells make adhesion only to the membrane itself,
though on the opposite sides.
This method resemble more the in vivo situation, and allows multiple
analyses on both cell types. The presence of the membrane between
the two cell types and the complete separation of their own
supernatants permit to perform permeability assays and separate
assays in the two compartments, and to challenge one cell and then
measure the effects on the other cell type. Further, cells on both
sides of the membrane can be immunostained for any molecule of
interest, and analysed by light microscopy, electron microscopy and
transmission microscopy, allowing direct evaluation of cell protein
expression. Compared to artificial capillaries, our system is cheaper,
easier to use, and does not require particular equipment.
Stage of development.
We are presently using the system to test the importance of neuronlike signaling between podocytes and endothelial cells.
The co-culture system could be extremely useful to evaluate
alteration of permeability of the filtering barrier in different conditions.
A better understanding of the processes at the basis of glomerular
damage and of the physiological properties of the glomerular filtration
barrier are mandatory to improve diagnosis and find more targeted
therapies. Besides the evident help to basic research studies, we
believe that our device can help in discovering and testing potential
novel biomarkers, individual patient samples, and the effects of new
drugs.
Fondazione seeks a commercial partner interested in the
acquisition of new disease models to be exploited for the
identification of new therapeutic agents and/or for the
development of diagnostic/prognostic tools.
Technology offer
□ A new 3D co-culture method of podocytes
and endothelial cells.
Inventor
Rastaldi Maria Pia and Li Min
Applications
□ Morphological and functional assays.
□ In vitro model for pathologies.
□Cellular toxicity assay for drugs.
Novel aspects and advantages
□Effective 3D method for culturing podocytes
and endothelial cells.
□
□ Cheaper and easier to use.
Current development stage
Applied basic research.
Intellectual property rights
□ Italian Patent application MI2009A00221617
□ International Patent application
PCT/IB20110/003298
Offer
□ collaboration for product development
□ patent licensing