Isolation of Viable Adipocytes and Stromal Vascular Fraction from Human Visceral Adipose Tissue Suitable for RNA Analysis and Macrophage PhenotypingGuadalupe Estrada-Gutierrez 1, Eyerahi Bravo-Flores 2, Veronica Ortega-Castillo 3, Veronica Flores-Rueda 4, Ismael Mancilla-Herrera 5, Salvador Espino Y Sosa 6, Maribel Sánchez-Martínez 2, Otilia Perichart-Perera 7, Mario Solis-Paredes 8
1Research Division, Instituto Nacional de Perinatologia, 2Department of Immunobiochemistry, Instituto Nacional de Perinatologia, 3Department of Obstetrics, Instituto Nacional de Perinatologia, 4Department of Academic Programs and Continuing Education, Instituto Nacional de Perinatologia, 5Department of Infectology and Immunology, Instituto Nacional de Perinatologia, 6Clinical Research Branch, Instituto Nacional de Perinatologia, 7Department of Nutrition and Bioprogramming, Instituto Nacional de Perinatologia, 8Department of Human Genetics and Genomics, Instituto Nacional de Perinatologia, Mexico City, Mexico
This protocol provides an efficient collagenase digestion method for isolation of viable adipocytes and stromal vascular fraction-SVF cells from human visceral fat in a single process, including methodology to obtain high-quality RNA from adipocytes and phenotypification of SVF-macrophages through staining of multiple membrane-bound markers for analysis by flow cytometry.
