Facilities and Equipment for Research

The Biochemistry and Molecular Biology laboratory is set up for the preparation of biological samples with individual activity benches, a chemical hood for the safe handling of toxic reagents, a cabinet dedicated to dangerous reagents/substances, centrifuges suitable for different activities, agitators, thermostats, a balance, a pH meter, refrigerators +4°C and freezers -20°C for the storage of reagents. Finally, there is also the Automated Liquid Handling Workstation – Microlab Starlet 496 for the automatic and precise preparation of large numbers of biological samples for both nucleic acid and protein extraction. As far as analytical activities are concerned, there is a spectrophotometer (Nanodrop) for the quantification of biological extracts, a multi-mode plate reader (Synergy 2 – BioTek) for fluorescence, absorbance and luminescence analysis, electrophoretic cells for agarose gel travel of proteins and nucleic acids, a Gel Doc Xr (Bio-RaD) and a C-Digit blot.

The laboratory is equipped with all the necessary equipment for the development and execution of cell and molecular biology experiments. The presence of three laminar flow hoods and two incubators ensures the correct management of primary and line cell cultures; in addition, a hood is specially dedicated to the process of blood samples and biopsy derived cells. The number of counters ensures a correct distribution of space for conducting experiments in biochemistry, cell biology and molecular biology. The laboratory is also equipped with two electrophoresis systems, both single and two-dimensional, for the rapid transfer (7 minutes) of proteins from gel to membrane, and subsequent analysis by dedicated scanner. In addition, the RANDOX IMMUNOASSAY system is available for multi-parameter cytokine analysis from sera and plasma. Finally, the laboratory is equipped with the MG140/200 incubator with automatic egg vault for conducting the corio-allantoid membrane test (CAM) and a thermal cycler for Real-Time PCR, connected to a PC equipped with special software for the execution of experiments, the visualization of results and the analysis of raw data.

The laboratory is dedicated to cell cultures of both line (i. e. endothelial cells) and primary (hematic and tissue samples of both healthy and pathological patients). In addition, all functional in vitro analyses and assays for the evaluation of proliferation, vitality and migration relevant to the planned studies are performed. These activities are made possible by the presence of two laminar flow hoods, three incubators, a thermostated bath and a refrigerated centrifuge.

In the BL2 laboratory, structured according to the standards established by the Authorities responsible, Genetically Modified Virus viruses (class 2) are manipulated and used, both for the modulation of specific genes and for reprogramming aimed at the production of iPS cells by viral infection and the subsequent differentiation towards functional cardiomyocytes, differentiation obtained also from embryonic stem cells. Given the high standards of biological safety, the room is also used for the handling and differentiation, specifically towards the cardiomyocyte line, of stem cells.

They are present in the laboratory: an cytofluorimeter (FACS Canto II BD – 3 laser configuration) for phenotypic characterisation and/or multi-parameter, qualitative and quantitative analysis of specific biomarkers; a cell-sorter (FACS Aria II BD – configuration with 3 lasers) that, in addition to allowing the cytofluorimetric analysis, allows, starting from a heterogeneous population, to physically separate and collect specific subpopulations according to the antigenic expression of interest. In particular, with these instruments lymphocyte subpopulations (T, B and NK lymphocytes), circulating endothelial progenitor cells (Endothelial Progenitor Cells – EPCs), circulating endothelial cells obtained from umbilical cord vein (HUVEC), pericides and multipotential stem cells are analyzed in the following areas.

The laboratory is equipped with all the necessary equipment for the activities of: cell culture (laminar flow cup, CO2-controlled incubators, thermostat bath, refrigerated centrifuge, fluorescence microscope and automatic system for cell count and molecular biology (chemical cape, electrophoresis system, centrifugal and microcentrifuge chilled). Finally, there is a freezer -20°C and a refrigerator + 4°C for the storage of reagents. The laboratory is mainly dedicated to studies on diabetes and dysmetabolic diseases.

The laboratory is equipped with all the equipment for both cell culture and molecular biology activities. There is therefore a laminar flow hood, two chemical hoods, a thermostat bath, a CO2-controlled incubator, a refrigerated centrifuge, an optical microscope, a refrigerator +4°C and a freezer -20°C, a thermal cycler, a thermomixer, a vortex, an orbital shaker and a pHmeter.

The area adjacent to the warehouse is equipped with: – a cold chamber: it allows to store at a controlled temperature +4° consumption material, such as soil and supplements for cell culture, biochemical analysis kit and cell biology. two liquid nitrogen tanks for the preservation of cells and tissues. a refrigerator-freezer park -20°, +4°, -80° for the storage and conservation of both consumables for experiments and biological samples. A cryopreservation facility is also available for the long-term storage of biological material (cellular and primary lines, stem cells, tissues, biological liquids, nucleic acids) at very low temperatures: liquid nitrogen (-196°C), nitrogen vapours (-140°C) and mechanical freezers (-80°C). The facility includes an integrated h24 SCUBE operational operational monitoring and control system capable of monitoring all critical equipment in the facility together with a latest generation of alarm and remote storage system. Each individual sample is recorded in the central database and labelled with a two-dimensional barcode. This highly advanced database maintains a complete history of each sample, so that individual events can be traced: when a sample has been received, defrosted, fractionated, tested or manipulated.

The following activities are carried out in the laboratory: – Extraction of RNA and proteins from cell samples; – Colorations of immunofluorescence on fixed cells and slides with slices of paraphinized tissue. The laboratory is also equipped with a transfer blots and freezer -80°C for storage of cellular extracts.

Microbiology experiments are carried out in the laboratory (thermostatted agitator and laminar flow hood), preparation of solutions with harmful reagents (chemical cape), electrophoretic strokes and fractionation of nucleic acids and proteins (horizontal and vertical electrophoretic cells with the appropriate feeders).