Last two decades, mSKIL has developed innovative and elegant in vitro models using primary cells for studying musculoskeletal and rheumatic diseases. We have in vitro models and technologies for studying cytokines network, oxidative stress, senescence, cartilage repair and degradation, bone remodeling, synovial inflammation and skeletal muscle cells metabolism. We are expert in co-culture systems and in the culture of cells under mechanical strains (Flexcell system). Each model is conceptualized to be as close as possible to the real in vivo condition, sometimes using 3D technologies. ![]() The lab has a specific room dedicated to mammalian cell-culture with two Class II laminar flow hoods, one humid 37 degree CO2 incubator, one water bath and one centrifuge. It also has one Zeiss inverted microscope coupled with high-definition camera, one autoclave, several refrigerators, freezers and one minus 80 degree ultrafreezer. In the lab and also with its full access to GIGA and Center for Interdisciplinary Research On Medicines (CIRM) platforms, mSKIL can perform all up-to-date imaging and histology technologies, even up to electronic microscopy. Recently, we use the Nanolive CX-A equipment to aquire refractive index live cell imaging. Up to 72h time-lapse of cell differentiation can be record, without any fixative or staining biais. Classical fluorochrome can also be add (throught FITC, TRICT, DAPI and Cy5 channel). ![]() The lab has a Leica semi-automatic microtome and a Zeiss Axiolab microscope with professional objectives coupled with a high-definition color camera to capture the best of our paraffin stained slides. We have also a dedicated fume-hood to perform all classical, specific stainings (Alcian blue, toluidine blue, safranin-O fast green, Movat's pentachrome...) or immunohistochemistry. We also collaborate with Rowiak to access to the laser microtome sectioning on every tissue or biomaterial to avoid artefact in very hard tissue/material. Immunofluorescence experiment on cells or tissues is routinely performed in the lab and capture in the GIGA imaging platform. We can investigate the whole transcriptomic activity of compounds, using RNA sequencing technology (RNAseq) or classical RT-qPCR, in either primary chondrocytes, osteoblasts, synoviocytes or myoblasts coming from healthy or diseased patients. We can perform these studies using our different innovative in vitro models, as close as possible to the real in vivo context. ![]() Single cell gene expression experiment is also possible. Data analysis is then made by mSKIL's scientists in R software using DESeq2, edgeR, limma or other adequate statistical package for differential gene expression profiling. Classical RT-qPCR is performed in the lab with a thermocycler Hybaid and a real-time PCR Rotor Gene device from Qiagen. The secretome, the global group of secreted proteins into the extracellular space by a cell at any given time and conditions, could be investigated in the lab by different methods. As we developed several ELISA biomarkers, we have a great expertise in immunoassays, from immunization to development and validation, with conventional mouse, rat or rabbit IgG to nanobodies. Western-blotting can be done in complementary experiments. We are also familiar with mass spectrometry in cell culture supernatant (LC MS/MS,...) that can be done in GIGA or in CIRM proteomic platforms. ![]() ELISA lab's equipment : one microplate washer (TECAN Hydroflex), two microplate shakers (Heidolph Titramax100), one spectrophotometer (THERMO Multiskan GO) and one fluorimeter (THERMO ascent). Thanks to a collaboration with mSKIL spin-off Artialis, multiplex immunoassays with high dynamic range are feasible using MesoScaleDiscovery (MSD) platform. Western-blotting is made using BioRad : Stain-Free Technology, Trans-Blot Turbo Transfer System and read with ChemiDoc MP. The technique, which allows the analysis of blood cells, cells isolated from tissue or from a cell line is an essential tool for very accurate cell phenotyping. Cell sorting or just count, with the simultaneous detection of several molecules, cell proliferation, DNA damage or cell viability. ![]() Cells are prepared in the lab and routinely analyzed in the GIGA flow cytometry platform by mSKIL's scientists. CatWalk XT from Noldus is a complete gait analysis system for quantitative assessment of footfalls and locomotion in mice, rats and guinea pigs. Illuminated Footprints allow detection of real footprints and body weight distribution. ![]() mSKIL has a special room, next to the animal facilities, dedicate to the CatWalk XT system. Thanks to our collaboration with our spin-off Artialis, we also has access to Smalgo, Dynamic Weight Bearing (DWB) and electronic von Frey. Among its many applications the Mach-1 system (Biomomentum) is used to assess the cartilage biomechanical properties. This equipment is able to perform indentation test, tension, shear and compression tests and mechanical stimulation on articular cartilage. This technique provides a complete characterization of cartilage mechanical properties and 3D mapping based on cartilage properties. This technique can be applied to other tissues, biomaterials or cells. ![]()
mSKIL has a special room dedicate to the Mach-1 device and all his accessories.
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