University of ferrara

6 ARTICLES PUBLISHED IN JoVE

Bioengineering

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

Manuela Ferracin¹, Irene Salamon², Laura Lupini², Elena Miotto², Silvia Sabbioni³, Massimo Negrini⁴

¹Department of Experimental, Diagnostic and Specialty Medicine - DIMES, University of Bologna, ²Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, ³Department of Life Sciences and Biotechnology, University of Ferrara, ⁴Department of Morphology, Surgery and Experimental Medicine and Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara

A sensitive and accurate method for cell-free microRNAs quantification using a dye-based chemistry and droplet digital PCR technology is described.

Neuroscience

Personalized Needles for Microinjections in the Rodent Brain

Giovanna Paolone^*1, Chiara Falcicchia^*1, Gianluca Verlengia^*1, Mario Barbieri¹, Anna Binaschi¹, Federico Paliotto¹, Beatrice Paradiso¹, Marie Soukupova¹, Silvia Zucchini^1,2, Michele Simonato^1,2

¹Department of Medical Sciences, Section of Pharmacology, and National Institute of Neurosciences, University of Ferrara, ²Laboratory for the Technologies for Advanced Therapies (LTTA), University of Ferrara

We describe here a protocol for microinjection in the rodent brain that uses quartz needles. These needles do not produce detectable tissue damage and ensure reliable delivery even in deep regions. Moreover, they can be adapted to research needs by personalized designs and can be re-used.

Neuroscience

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Marie Soukupová¹, Chiara Falcicchia¹, Francesca Lovisari¹, Selene Ingusci¹, Mario Barbieri¹, Silvia Zucchini¹, Michele Simonato¹

¹Department of Medical Sciences, Section of Pharmacology, Neuroscience Center, University of Ferrara and National Institute of Neuroscience

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

Biology

Human Adipose Tissue Micro-fragmentation for Cell Phenotyping and Secretome Characterization

Bianca Vezzani^1,2, Mario Gomez-Salazar¹, Joan Casamitjana¹, Carlo Tremolada³, Bruno Péault^1,4

¹MRC Center for Regenerative Medicine, University of Edinburgh, ²Dept. of Morphology, Surgery and Experimental Medicine, Section of General Pathology, University of Ferrara, ³Italian Image Institute, ⁴Orthopaedic Hospital Research Center and Broad Stem Cell Research Center, David Geffen School of Medicine, University of California

Here, we present human adipose tissue enzyme-free micro-fragmentation using a closed system device. This new method allows the obtainment of sub-millimeter clusters of adipose tissue suitable for in vivo transplantation, in vitro culture, and further cell isolation and characterization.

JoVE Core

Cultivating a Three-dimensional Reconstructed Human Epidermis at a Large Scale

Irini M. Dijkhoff¹, Benedetta Petracca^1,2, Roxane Prieux³, Giuseppe Valacchi^3,4, Barbara Rothen-Rutishauser¹, Marc Eeman²

¹Adolphe Merkle Institute, University of Fribourg, ²Dow Silicones Belgium SRL, ³Department of Biomedical and Specialist Surgical Sciences, University of Ferrara, ⁴Department of Animal Sciences, Plants for Human Health Institute, North Carolina State University

This protocol describes a straightforward method to cultivate three-dimensional reconstituted human epidermis in a reproducible and robust manner. Additionally, it characterizes the structure-function relationship of the epidermal barrier model. The biological responses of the reconstituted human epidermis upon proinflammatory stimuli are also presented.

Medicine

Functional Characterization of Endogenously Expressed Human RYR1 Variants

Susan Treves^1,2, Thierry Girard³, Francesco Zorzato^1,2

¹Department of Biomedicine, Basel University Hospital, ²Department of Life Sciences, University of Ferrara, ³Department of Anesthesia, Basel University Hospital

Here methods used to study the functional effect of RYR1 mutations endogenously expressed in Epstein Barr Virus immortalized human B-lymphocytes and muscle biopsy derived satellite cells differentiated into myotubes are described.