STAR Protocols (Dec 2024)
Protocol for isolating CD163+ Kupffer cells from human liver resections
- Armando Andres Roca Suarez,
- Marie-Laure Plissonnier,
- Maud Michelet,
- Anaëlle Dubois,
- Sarah Heintz,
- Maria Saez-Palma,
- Marion Delphin,
- Isabelle Bordes,
- Jennifer Molle,
- Audrey Diederichs,
- Mélanie Rodà,
- Emmanuel Combe,
- Léa Mougené,
- Guillaume Giraud,
- Xavier Grand,
- Michel Rivoire,
- Massimo Levrero,
- Barbara Testoni,
- Fabien Zoulim
Affiliations
- Armando Andres Roca Suarez
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France; Corresponding author
- Marie-Laure Plissonnier
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Maud Michelet
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Anaëlle Dubois
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Sarah Heintz
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Maria Saez-Palma
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Marion Delphin
- The Francis Crick Institute, London, UK
- Isabelle Bordes
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Jennifer Molle
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Audrey Diederichs
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Mélanie Rodà
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Emmanuel Combe
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Léa Mougené
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Guillaume Giraud
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Xavier Grand
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France
- Michel Rivoire
- The Lyon Hepatology Institute EVEREST, Lyon, France; INSERM U1032, Centre Léon Bérard (CLB), 69008 Lyon, France
- Massimo Levrero
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France; Department of Hepatology, Croix Rousse Hospital, Hospices Civils de Lyon, Lyon, France; Department of Internal Medicine - DMISM and the IIT Center for Life Nanoscience (CLNS), Sapienza University, Rome, Italy
- Barbara Testoni
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France; Corresponding author
- Fabien Zoulim
- INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France; Department of Hepatology, Croix Rousse Hospital, Hospices Civils de Lyon, Lyon, France; Corresponding author
- Journal volume & issue
-
Vol. 5,
no. 4
p. 103359
Abstract
Summary: The liver microenvironment contains a wide variety of monocyte and macrophage populations. Here, we present a protocol for the specific isolation of liver-resident macrophages, known as Kupffer cells (KCs), from human liver resections. We describe steps for dissociating human liver tissues, separating non-parenchymal cells into fractions by a 2-phase iodixanol gradient, and positive selection of KCs based on the expression of CD163. We then provide instructions for validating the procedure by immunofluorescence to detect CD163.For complete details on the use and execution of this protocol, please refer to Roca Suarez, Plissonnier, et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.