Laboratory of Experimental Pharmacology of Endothelium (JCET)

The research group led by Professor Stefan Chłopicki has many years of experience in research on the mechanisms of endothelial dysfunction in atherosclerosis, heart failure and other diseases, and on vasoprotective mechanisms of endothelium that could be therapeutically targeted by first-in-class small molecule PGI2 releasers, CO-donors or many others. The up allowing studies on vessels from various vascular beds; conductive as well as resistance vessels) and isolated perfused organs preparations (isolated lungs, isolated heart, isolated liver, isolated kidney). Studies in the laboratory involve multiple animal models of endothelial dysfunction (e.g. heart failure, atherosclerosis, pulmonary hypertension, hipertrigliceridemia, diabetes or inflammatory liver diseases) In various in vitro, ex vivo and in vivo models listed above, the activity of the protective (e.g. NO, PGI2), proinflammatory (e.g. MCP-1, IL-6), prothrombotic (e.g. TM) endothelial mediators is analyzed, which reflects the functional endothelial phenotype and the efficacy of endothelium-targeted therapy. An important supplement in the studies performed in the laboratory is the assessment of morphological and biochemical blood parameters as well as histological and biochemical parameters of the vessel wall. The laboratory develops comprehensive in vivo pharmacology of endothelium in multiple disease models, for better understanding of the therapeutic potential of endothelial and vascular wall mechanisms targeted by various therapeutics.

An 8-channel Tissue Organ Bath System (Danish Myo Technology – DMT) with an advanced semi-automatic washing system designed for studies of isolated blood vessels of larger
diameter (2 mm). A set of two 4-channel Myograph Systems (DMT) for studying isolated blood vessel rings of smaller diameter (eg. mice aorta of 1-2 mm diameter or mesenteric
artery of approx. 100 μm diameter).

Two Pressure Myograph Systems (DMT) for the studies of the small cannulated blood vessel(diameter 60-500 μM) under flow condition (through vessel lumen) – close to the
physiological situation.

Pressure Myograph Culture Systems (DMT) including 4 channel set-up for prolonged studies of the cannulated blood vessel activity (of the diameter from 60 μm to 2 mm).

Isolated Perfused Ventilated Lungs (HSE-HA) allowing the studies of isolated rat or mice lungs ex vivo close to physiological conditions. Isolated lungs placed in an artificial thorax are ventilated with negative pressure. The equipment allows the simultaneous assessment of respiratory activity parameters (e.g.: tidal volume, respiratory flow, lung volume), the parameters of pulmonary circulation (e.g.: pulmonary arterial and venous pressure) and pulmonary edema of isolated lungs. The apparatus allows the perfusion of the pulmonary circulation under constant pressure or constant flow conditions.

Isolated Working Heart System (Hugo Sachs Elektronic-Harvard Apparatus, HSE-HA) allowing the studies of the working mouse (or rat) heart activity ex vivo, where the heart is physiologically perfused through its left atrium and left ventricle.

Mice Treadmill with Oxymax Chamber (Columbus Instruments). The apparatus is dedicated to measure the animal oxygen consumption and excretion of carbon dioxide (CO2) during controlled physical activity of mouse on the treadmill and allows the measurement of the exercise capacity (VO2 max).

ApoGee A50-Micro (ApoGee Flow Systems Ltd, Hemel Hempstead, UK) – high resolution flow cytometer with two lasers, three light scatter detectors and six fluorescence detectors designed for microvesicles analysis.

Izon qNano (Izon Science Ltd, Christchurch, New Zealand) – instrument using tunable resistive pulse sensing technique for direct measurement of microvesicles size, diameter and concentration.

Biochemical room

ABX Pentra 400 biochemical analyzer (Horiba Medical) allows automated measurement of 55 basic serum or plasma parameters by spectrophotometric or potentiometric methods .

pHOx Ultra Analyzer (Nova Biomedical) is designed to measure blood gasometric parameters (PO2, PCO2), to perform complete oximetry tests (i.e. measurement of O2-Hb, CO-Hb, Met-Hb) and to measure basic blood electrolytes.

Histology room

Electromechanical cryostat with a system for keeping the chamber clean and tidy (Leica CM 1950), manual rotation microtome (Sakura Accu-Cut SRM), paraffin embedding centre (Thermo Scientific Microm EC 350-2), benchtop tissue processor (Leica TP 1020), apparatus for automatic sample staining (Leica Autostainer XL).

Axio Imager.A2 Fluorescence Microscope (Carl Zeiss) and Axio Observer.D1 inverted fluorescence microscope (Carl Zeiss) with filter set (set of filters) for three spectral channels, and the ability to image with DIC interference contrast and PlasDIC; equipped with monochromatic and multicolor cameras.

Virtual microscopy centre based on the automated Olympus BX51 microscope for scanning and archiving histological samples as high resolution images. The scanner software allows online image sharing (virtual histology).

ScanR screening system consisting of fully automated inverted fluorescence microscope (Olympus IX81), placed in an incubator with a system allowing long term microenvironment maintenance, appropriate for cell culturing (Life Imaging Services). The whole system offers a unique combination of a fluorescence microscope with an image cytometer, allowing real-time cell phenotype analysis under physiological conditions.