Accedi

sapienza university

2 ARTICLES PUBLISHED IN JoVE

Biology

Isolation, Culture, and Functional Characterization of Adult Mouse Cardiomyoctyes

Evan Lee Graham¹, Cristina Balla^1,2, Hannabeth Franchino¹, Yonathan Melman¹, 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.

Medicine

Inducing and Characterizing Vesicular Steatosis in Differentiated HepaRG Cells

Silvia Di Cocco^1,2, Laura Belloni³, Abigail D.G. Nunn³, Debora Salerno³, 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.