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sapienza university

2 ARTICLES PUBLISHED IN JoVE

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Biology

Isolation, Culture, and Functional Characterization of Adult Mouse Cardiomyoctyes
Evan Lee Graham 1, Cristina Balla 1,2, Hannabeth Franchino 1, Yonathan Melman 1, Federica del Monte *1, Saumya Das *1
1Cardiovascular Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, 2Division of Cardiology, Sapienza University

Here we describe the isolation of adult mouse cardiomyoctyes using a Langendorff perfusion system. The resulting cells are Ca2+-tolerant, electrically quiescent and can be cultured and transfected with adeno- or lentiviruses to manipulate gene expression. Their functionality can also be analyzed using the MMSYS system and patch clamp techniques.

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Medicine

Inducing and Characterizing Vesicular Steatosis in Differentiated HepaRG Cells
Silvia Di Cocco 1,2, Laura Belloni 3, Abigail D.G. Nunn 3, Debora Salerno 3, Silvia Piconese 2,4, Massimo Levrero 2,3,5,6, Natalia Pediconi 1,3
1Department of Molecular Medicine, Sapienza University, 2Department of Internal Medicine - DMISM, Sapienza University, 3Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, 4Pasteur Institute Italy-Fondazione Cenci Bolognetti, 5INSERM U1052-Cancer Research Center of Lyon, 6Department of Hepatology, Croix Rousse Hospital, Hospices Civils de Lyon

In this study, we describe a detailed protocol for inducing liver vesicular steatosis in differentiated HepaRG cells with the fatty acid salt sodium oleate and employ methods for detection and quantification of lipid accumulation, including coherent anti-Stokes Raman scattering (CARS) microscopy, cytofluorimetric analysis, Oil red O staining, and qPCR. 

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