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6 ARTICLES PUBLISHED IN JoVE

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Medicine

Three-Dimensional (3D) Tumor Spheroid Invasion Assay
Maria Vinci 1,2, Carol Box 2, Suzanne A. Eccles 2
1Division of Molecular Pathology, The Institute of Cancer Research, 2Division of Cancer Therapeutics, The Institute of Cancer Research

Invasion of surrounding normal tissues is a defining characteristic of malignant tumors. We provide here a simple, semi-automated micro-plate assay of invasion into a natural 3D biomatrix that has been exemplified with a number of models of advanced human cancers.

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Cancer Research

Development of a Human Preclinical Model of Osteoclastogenesis from Peripheral Blood Monocytes Co-cultured with Breast Cancer Cell Lines
Laura Mercatali 1, Chiara Spadazzi 1, Giacomo Miserocchi 1, Chiara Liverani 1, Alessandro De Vita 1, Alberto Bongiovanni 1, Federica Recine 1, Dino Amadori 1, Toni Ibrahim 1
1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), IRCCS

This protocol describes development of an in vitro human preclinical model of osteoclastogenesis from peripheral blood monocytes cultured with breast cancer cell lines to mimic the cancer cell-osteoclast interaction. The model could be used to further our understanding of bone metastasis formation and improve therapeutic options.

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Cancer Research

Establishment of a Primary Culture of Patient-derived Soft Tissue Sarcoma
Alessandro De Vita 1, Laura Mercatali 1, Giacomo Miserocchi 1, Chiara Liverani 1, Chiara Spadazzi 1, Federica Recine 1, Alberto Bongiovanni 1, Federica Pieri 2, Davide Cavaliere 3, Valentina Fausti 1, Dino Amadori 1, Toni Ibrahim 1
1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Pathology Unit, Morgagni-Pierantoni Hospital, 3Unit of Surgery and Advanced Oncologic Therapies, Morgagni-Pierantoni Hospital

The following protocol focuses on the establishment of a primary culture of patient-derived soft tissue sarcoma (STS). This model could help us to better understand the molecular background and pharmacological profile of these rare malignancies and could represent a starting point for further research aimed at improving STS management.

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Cancer Research

Dried Blood and Serum Spots As A Useful Tool for Sample Storage to Evaluate Cancer Biomarkers
Laura Mercatali 1, Patrizia Serra 2, Giacomo Miserocchi 1, Chiara Spadazzi 1, Chiara Liverani 1, Alessandro De Vita 1, Giorgia Marisi 3, Alberto Bongiovanni 1, Federica Recine 1, Akwilina Pangan 4, Nestory Masalu 4, Toni Ibrahim 1, Dino Amadori 1
1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 2Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 3Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, 4Department of Oncology, Bugando Medical Center, Mwanza, Tanzania

This protocol describes a simple and useful method to store peripheral blood and serum/plasma for downstream analyses such as single nucleotide polymorphism (SNP) evaluation and ELISA assay.

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Biology

Measuring the Confluence of iPSCs Using an Automated Imaging System
Valentina Magliocca 1,2, Maria Vinci 3, Tiziana Persichini 2, Franco Locatelli 3, Marco Tartaglia 1, Claudia Compagnucci 1
1Genetics and Rare Diseases Research Division, Ospedale Pediatrico Bambino Gesù, IRCCS, 2Department of Science, University Roma Tre, 3Department of Onco-hematology, Gene and Cell Therapy, Ospedale Pediatrico Bambino Gesù, IRCCS

The goal of the protocol is to compare different extracellular matrix (ECM) coating conditions to assess how differential coating affects the growth rate of induced pluripotent stem cells (iPSCs). In particular, we aim to set up conditions to obtain optimal growth of iPSC cultures.

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Cancer Research

Live-3D-Cell Immunocytochemistry Assays of Pediatric Diffuse Midline Glioma
Giulia Pericoli *1, Roberta Ferretti *1, Andrew S. Moore 2,3, Maria Vinci 1
1Department of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children's Hospital, IRCCS, 2Oncology Service, Children's Health Queensland Hospital and Health Service, 3Child Health Research Centre, The University of Queensland

This study presents a protocol of live-3D-cell immunocytochemistry applied to a pediatric diffuse midline glioma cell line, useful to study in real-time the expression of proteins on the plasma membrane during dynamic processes like 3D cell invasion and migration.

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