Clinical Academic Center of Coimbra (CACC)

2 ARTICLES PUBLISHED IN JoVE

Cancer Research

Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors

Mafalda Laranjo^*1,2,3,4, Maria João Carvalho^*1,2,3,4,5,6, Beatriz Serambeque^1,2,3, André Alves^2,7, Carlos Miguel Marto^1,2,3,4,8, Isabel Silva⁹, Artur Paiva^3,9,10, Maria Filomena Botelho^1,2,3,4,8

¹Institute of Biophysics, Faculty of Medicine, University of Coimbra, ²Coimbra Institute for Clinical and Biomedical Research (iCBR), area of Environment Genetics and Oncobiology (CIMAGO), Faculty of Medicine, University of Coimbra, ³CNC.IBILI/Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, ⁴Clinical Academic Center of Coimbra (CACC), ⁵Universitary Clinic of Gynecology, Faculty of Medicine, University of Coimbra, ⁶Gynecology A Service, Coimbra Hospital and Universitary Center, ⁷Institute of Pharmacology & Experimental Therapeutics, Faculty of Medicine, University of Coimbra, ⁸Institute of Experimental Pathology, Faculty of Medicine, University of Coimbra, ⁹Cytometry Operational Management Unit, Clinical Pathology Service, Coimbra Hospital and Universitary Center, ¹⁰Polytechnic Institute of Coimbra, ESTESC-Coimbra Health School, Laboratory Biomedical Sciences

The aim of this methodology is to identify cancer stem cells (CSC) in cancer cell lines and primary human tumor samples with the sphere-forming protocol, in a robust manner, using functional assays and phenotypic characterization with flow cytometry and Western blot.

Biochemistry

Analysis of Fluorescent-Stained Lipid Droplets with 3D Reconstruction for Hepatic Steatosis Assessment

Karina Garcia^1,2,3,4, André Alves^1,2,3, Teresa M. Ribeiro-Rodrigues^1,2,3, Flávio Reis^1,2,3, Sofia Viana^1,2,3,4

¹Institute of Pharmacology & Experimental Therapeutics & Coimbra Institute for Clinical and Biomedical Research (iCBR), Faculty of Medicine, University of Coimbra, ²Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, ³Clinical Academic Center of Coimbra (CACC), ⁴Polytechnic Institute of Coimbra, Coimbra Health School

Herein, we demonstrate an optimized BODIPY 493/503 fluorescence-based protocol for lipid droplet characterization in liver tissue. Through the use of orthogonal projections and 3D reconstructions, the fluorophore allows for successful discrimination between microvesicular and macrovesicular steatosis and may represent a complementary approach to the classical histological protocols for hepatic steatosis assessment.