The Sadayappan Lab in the Department of Internal Medicine at the University of Cincinnati, College of Medicine (UCCOM) is located in the Cardiovascular Center Building, Rooms 4970- 4978 and Rooms 3968-3974. The lab consists of over 4000 sq. ft. and has all the necessary space and equipment for molecular, biological and biochemical experiments, as well as histological and immunohistochemical analyses. Students and trainees have designated bench space to perform experiments with all the equipment required to complete their respective projects. Available in the PI’s lab are desktop Eppendorf centrifuges, PCR and real-time PCR machines, gel documentation system, a nanodrop, shaking culture incubators, complete setup for expressing and purifying recombinant proteins from bacteria, spectrophotometers, abiosafety cabinets, -80˚C freezers, a microplate ELISA reader, protein electrophoresis units, Western blotting apparatus, etc. All experiments follow safety measures, as established by NIH policies and the Institutional Biosafety Committee of the UCCOM.

The lab also has access to core facilities at UCCOM and Cincinnati Children’s Hospital Medical Center (CCHMC) for additional support on complex experiments. Dr. Sadayappan has a joint appointment with the Heart Institute at CCHMC.

Facilities located in the Sadayappan Lab :

» Calcium and Contractility in Isolated Myocytes : The lab studies the heart at the cellular level using isolated cardiac muscle cells or cardiomyocytes. The IonOptix system allows us to investigate cellular dimensioning and intracellular calcium as important measures of the excitation-contraction coupling.

» Muscle Mechanical Studies : The lab has the latest instruments to measure force development, tension cost, ATPase activity and myofilament calcium sensitivity in skinned cardiac (1) papillary muscles and (2) myocytes (Aurora Scientific Inc).

» IntactSkeletal Muscle Functional Assay : Complete set of Dual-Mode Muscle Lever system (Aurora Scientific Inc) is equipped in our lab to test isometric and isotonic functional capacities of intact skeletal muscle in vivo, in situ, and in vitro.

» Dissecting Microscopes : Numerous Nikon (SMZ-10) and Olympus (BX60) dissecting microscopes are available in the lab for dissecting cardiac muscles for biophysical experiments, as well as taking pictures of TTC-stained heart sections and whole organs post-ischemia- reperfusion injury.

» Fluorescent Microscope : The lab has an Olympus IX73 fluorescent microscope with camera attachment to aid in immunohistochemistry documentation.

» In vitro Motility Assay : The lab has access to Nikon A1R LUN-Vinverted confocal fluorescent microscope and the reagents required to measure the motility of actin filaments as a function of acto-myosin interaction, along with the rotation of fluorescent myosin rods as a function of mechanical stability in myosin coiled coil.

» Adult Cardiomyocyte Preparation : A Langendorff preparation unit (Radnoti, Monrovia, PA) is available to perfuse the adult mouse hearts and isolate adult cardiomyocytes.

» C02 Incubator and Culture Hoods : The PI has a two separate BSL 2 level cell culture facilities: approved by the Institutional Biosafety Committee. Each room is equipped with a biosafety cabinet, C02 incubators, 37˚C bead bath, refrigerator and inverted microscopes.

Shared facilities available to the Sadayappan lab :

https://med.uc.edu/research/core-facilities : The UC College of Medicine houses a number of research core facilities designated as core service centers.

https://www.cincinnatichildrens.org/research/cores : Cincinnati Children’s Research Foundation is dedicated to advancing basic, translational, clinical and outcomes-based research. The core provides cutting-edge, cost-effective technology and data analysis that would be unattainable on an individual research basis. The fee-for-service program also offers unique studies that you can’t find anywhere else.

» Animal Facilities : UCCOM has AAALAC accredited animal facilities, which meet, or exceed, all federal, state and local regulations. These facilities provide daily animal husbandry and care, and provide comprehensive veterinary medical programs to monitor and maintain the health and well-being of these animals. Veterinary care is provided by full-time Laboratory Animal Veterinarians. UCCOM animal facility has ~71,000 ft 2 facilities which contain surgical, radiologic, fluoroscopic, necropsy, quarantine and training facilities.

» In Vivo Cardiac Function : Echocardiography equipment is available to the lab for measuring cardiac performance using the VisualSonics Vevo 2100 Imaging System (Canada).

» Mouse Physiology Core : This facility is for the measurement of cardiac physiological variables in the intact (often conscious) transgenic and knockout mouse. The Mouse Physiology Core Facility provides the additional laboratory equipment, instrumentation and technical help to evaluate sophisticated aspects of cardiovascular performance in transgenic and knockout mice.

» Gene Targeting / Transgenic Mouse Core : This core presents an exceptional opportunity for trainees and has facilities and resources for the production and characterization of transgenic and knockout mice. The facility can provide design expertise or manpower for targeting vector construction, and it introduces transgenes by pronuclear injection or homologous recombination in ES cells.

» Live Microscopy Core : The Live Microscopy Core supplies equipment and expertise that allow trainees to image living tissues with high-resolution light microscopy. The facility also provides access to, training in, and use of Metamorph software for image analysis. In addition, Flow Cytometry and Cell Sorting Core offers cell sorting and analysis capability and expertise to the entire clinical and research community at UCCOM.

» Morphology Core : This core includes technical support, cryostat, microtomes, embedding table, histochemical, immunohistochemical and radiographic supplies to perform routine staining, in situ autoradiography, and electron microscopy.

» Mass Spectrometry Facility : Recently, the State of Ohio Mass Spectrometry Consortium was formed through a partnership of 10 Ohio universities. The University of Cincinnati Mass Spectrometry Facility is one of the two primary centers in the Consortium. The Facility has recently acquired several state-of- the-art instruments that enable rapid screening of samples, trace metal analysis, and detailed structural analysis and characterization of biological molecules and organic and inorganic materials.

» Vector Production Facility (VPF) : The VPF is used for aseptic manufacture of early-phase GMP-compliant products, including, but not limited to, Retroviral, Adeno-associated Virus (AAV), Adenovirus (Ad) and Lentivirus vectors, Master Cell Banks, Master Viral Banks and other vector systems at BSL2.

» Bioinformatics Core : The Bioinformatics Core provides access to computer and software systems, database infrastructure, and direct user help for the characterization of gene or protein networks that exhibit coordinated regulation and cooperative function across the biological models under study. The Bioinformatics Core aids investigators in the design and analysis of complex, high-content experiments.

» Biostatistics Core, Center for Epidemiological and Biostatistics : This core provides consultation and/or ongoing support to researchers at UCCOM, such as development of clinical research plans, guidance and assistance in the development of grant proposals, data entry and management services using Teleform, and data analysis.

» Center for Stem Cell & Organoid Medicine (CuSTOM) : The CuSTOM at Cincinnati Children’s is a multi-disciplinary team of scientists, clinicians and entrepreneurs using advancements in developmental biology and stem cell technologies to revolutionize personalized medicine and improve patient care. The scientists in this forum have developed a way of using stem cells from patients to generate miniature organs known as organoids. These organoids are a revolutionary platform for understanding disease, developing new personalized treatments and, ultimately, generating tissue for transplantation.
https://www.cincinnatichildrens.org/research/divisions/c/CuSTOM