HIGLY QUALIFIED SERVICES FOR DRUG/BIOMATERIAL SCREENING
The lab focuses its research on angiogenesis and oncological pharmacology. It is particularly skilled in cellular biology, protein expression and enzyme activity and histological analysis, to test
treatment efficacy and safety. Flexibility, integration, technical capability, high scientific knowledge, constructive approach in problem solving and effective project management are some of the resources used to build strong, synergistic and successful partnerships.
In vitro cell proliferation and survival (price per test substance and per cell type: 750 Euro)
Culturing of human endothelial or tumor cells (or alternative cell line) in monolayer or in suspension
Dose-responses in at least 3 diluted concentrations (in triplicate, n=3 different experiments)
Positive and negative controls for reference
MTT assay at one fixed time or microscopic cell counting
Statistical analysis and written report
In vitro cell migration by scratch assay (price per test substance and per cell type 800 Euro)
Culturing of human endothelial or tumors cells (or alternative cell line)
Dose-responses of 3 concentrations (in triplicate, n=3)
Positive and negative control for reference
Recovery of wound area following scratch of confluent monolayer
Image analysis at different times
Statistical analysis and written report
In vitro cell chemotaxis by Transwell chamber (price per test substance and per cell type 800 Euro)
Culturing of human endothelial or tumor cells (or alternative cell line)
Dose-responses of 3 concentrations (in triplicate, n=3)
Positive and negative control for reference
Cell counts at the microscope
Statistical analysis and written report
Permeability assay (price per test substance 1500 Euro)
Culturing of human endothelial cells on transwell inserts
Fixed concentration of the substance (in triplicate, n=3)
Two incubation times
Positive and negative control of permeability
Check of morphology after stimulation
Statistical analysis and written report
In vitro angiogenesis assay on Matrigel (price per test substance 1500 Euro)
Culturing of human endothelial cells
Seeding on Matrigel 3D gels
Dose-responses of 3 concentrations (in triplicate, n=3)
Positive and negative control of angiogenesis
Image analysis of endothelial meshwork organization at different times
Statistical analysis and written report
Molecular Biology and protein/gene expression (price per test substance and per cell type
800 Euro)
Study of the mechanisms of action through the evaluation of the expression/activation of growth factors (HIF1alfa, VEGF), signalling systems and enzymatic cascades (ROS production, matrix metalloproteinases), production and organization of cytoskeletal proteins and extracellular matrix (laminin, fibronectin, collagen elastin, endogenous hyaluronic acid) on cell cultures and tissue samples or sections. Evaluation of endothelial cell markers: ICAM-CD54, PECAM-CD31 on cell cultures and tissue samples or sections. Evaluation of inflammatory markers and mediators (COX isoforms, PGE synthases, inducible NOS.
Measurement of soluble mediators (NO, PGE2, interleukins, etc..)
Culturing of human endothelial or tumor cells or alternative cells
Fixed concentration of the substance vs untreated cells (in triplicate, n=3)
Appropriate incubation time
Methods specific for each marker: RT-PCR/western blot/immunofluorescence/ biochemicalassays/staining of histological sections
Image analysis, statistical analysis and written report
In vitro modeling of pathological processes (price per each model 3000 Euro)
Development of in vitro models to simulate specific disease states for the evaluation of therapeutic compounds or bioactive substances.
Set up of tumor multicellular spheroids: 3D cultures of tumor cells in the presence of stromal/endothelial cells/immune cells to recapitulate the tumor microenvironment (TME).
Possible compartmentalization of the cellular components to evaluate specific cell-cell communication.
Fibrosis Models: Induction of fibrotic phenotypes using specific agents (e.g., TGF-beta, IL1beta). Evaluation focuses on myofibroblast activation and extracellular matrix (ECM) deposition (collagen, fibronectin).
Endothelial-to-Mesenchymal Transition (EndMT): Recapitulation of the transition from endothelial to mesenchymal phenotypes. Monitoring the loss of endothelial markers (CD31, VE-cadherin) and the acquisition of mesenchymal markers (alpha-SMA, Vimentin).
Inflammation Models: Induction of inflammatory responses using specific stimuli (e.g., LPS, TNF-alpha, IL-1beta). Evaluation of cytokine release, inflammatory mediators (COX-2, iNOS), and signaling pathways.
Genetic Manipulation Services: genetic engineering to explore the functional relationship between specific genes and cellular phenotypes.
-Gene Silencing (Loss of Function) (price per gene 2500 Euro)
siRNA (Small Interfering RNA): For transient silencing and short-term studies.
shRNA (Short Hairpin RNA): For stable, long-term gene knockdown via viral vector delivery or plasmid transfection.
Validation of silencing
-Overexpression (Gain of Function) (price per gene 2500 Euro)
Plasmid Transfection/Viral Transduction: Introduction of exogenous DNA to increase the expression of specific genes, allowing for the study of protein overactivity or rescue experiments.
Validation of overexpression
Contact persons:
Prof. Lucia Morbidelli (lucia.mrobidelli@unsii.it)
Prof. Sandra Donnini (sandra.donnini@unisi.it)
Dr Valerio Ciccone (valerio.ciccone@unisi.it)