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Vita-Salute San Raffaele University

5 ARTICLES PUBLISHED IN JoVE

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JoVE Journal

Exploiting Live Imaging to Track Nuclei During Myoblast Differentiation and Fusion
Giorgia Careccia *1,2, Federica Colombo *1,3, Mario Tirone 1, Alessandra Agresti 1, Marco E. Bianchi 1,2, Samuel Zambrano *1,2, Emilie Vénéreau *1
1Chromatin Dynamics Unit, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Raffaele Scientific Institute, 2Vita-Salute San Raffaele University, 3Department of Electronics, Information and Bioengineering, Politecnico di Milano

Skeletal muscle differentiation is a highly dynamic process, which particularly relies on nuclear positioning. Here, we describe a method to track nuclei movements by live cell imaging during myoblast differentiation and myotube formation and to perform a quantitative characterization of nuclei dynamics by extracting information from automatic tracking.

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Immunology and Infection

Purification and Expansion of Mouse Invariant Natural Killer T Cells for in vitro and in vivo Studies
Gloria Delfanti 1, Alessandra Perini *1,2, Elisa Zappa *1,2, Maya Fedeli 1,2
1Experimental Immunology Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, 2Vita-Salute San Raffaele University

We describe a rapid and robust protocol to enrich invariant natural killer T (iNKT) cells from mouse spleen and expand them in vitro to suitable numbers for in vitro and in vivo studies.

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Medicine

Fertility Sparing Procedure using Carbon Dioxide Fiber Laser Vaporization of Ovarian Endometrioma
Massimo Candiani 1, Jessica Ottolina 1, Iacopo Tandoi 1, Ludovica Bartiromo 1, Matteo Schimberni 1, Roberta Villanacci 1, Stefano Ferrari 1
1Gynecological Surgery and Endometriosis Departmental Unit, San Raffaele Scientific Institute

In this protocol, CO2 fiber laser technique is demonstrated for the surgical treatment of ovarian endometriosis, which represents a viable alternative in terms of fertility preservation with the major advantage of not being dependent on the surgeon's skills and personal experience.

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Medicine

Laparoscopic Oocyte Retrieval and Cryopreservation during Vaginoplasty for Treatment of Mayer-Rokitansky-Kuster-Hauser Syndrome
Valeria Stella Vanni *1,2, Alessandra Alteri *1,2, Lucia De Santis 1,2, Greta Chiara Cermisoni 1, Elisa Rabellotti 1, Diana Delprato 1, Marta Parma 1, Enrico Papaleo 1, Luigi Fedele 3, Massimo Candiani 1,2
1Obstetrics and Gynaecology Unit, IRCCS San Raffaele Scientific Institute, 2Vita-Salute San Raffaele University, 3Department of Obstetrics and Gynecology, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico

A dedicated team can offer Mayer-Rokitansky-Kuster-Hauser patients the option to perform controlled ovarian stimulation and oocyte cryopreservation at the time of laparoscopic vaginoplasty.

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Bioengineering

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
Alessandro Migliara 1, Martino Alfredo Cappelluti 1, Francesca Giannese 2, Sara Valsoni 1, Alberto Coglot 1, Ivan Merelli 1,3, Davide Cittaro 2, Angelo Lombardo 1,4
1San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), IRCCS San Raffaele Scientific Institute, 2Center for Omics Sciences, IRCCS San Raffaele Institute, 3Institute for Biomedical Technologies, National Research Council, 4Vita-Salute San Raffaele University

Here, we present a protocol for the in vitro selection of engineered transcriptional repressors (ETRs) with high, long-term, stable, on-target silencing efficiency and low genome-wide, off-target activity. This workflow allows for reducing an initial, complex repertoire of candidate ETRs to a short list, suitable for further evaluation in therapeutically relevant settings.

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