STAR Protocols (Sep 2021)
Protocols for endothelial cell isolation from mouse tissues: kidney, spleen, and testis
- Sébastien J. Dumas,
- Elda Meta,
- Nadine V. Conchinha,
- Liliana Sokol,
- Rongyuan Chen,
- Mila Borri,
- Laure-Anne Teuwen,
- Koen Veys,
- Melissa García-Caballero,
- Vincent Geldhof,
- Lucas Treps,
- Pauline de Zeeuw,
- Kim D. Falkenberg,
- Charlotte Dubois,
- Magdalena Parys,
- Laura P.M.H. de Rooij,
- Katerina Rohlenova,
- Jermaine Goveia,
- Luc Schoonjans,
- Mieke Dewerchin,
- Guy Eelen,
- Xuri Li,
- Joanna Kalucka,
- Peter Carmeliet
Affiliations
- Sébastien J. Dumas
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Elda Meta
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Nadine V. Conchinha
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Liliana Sokol
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Rongyuan Chen
- State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong, P.R. China
- Mila Borri
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Laure-Anne Teuwen
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium; Translational Cancer Research Unit, GZA Hospitals Sint-Augustinus, Antwerp 2610, Belgium; Center for Oncological Research, University of Antwerp, Antwerp 2000, Belgium
- Koen Veys
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Melissa García-Caballero
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Vincent Geldhof
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Lucas Treps
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Pauline de Zeeuw
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Kim D. Falkenberg
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Charlotte Dubois
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Magdalena Parys
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Laura P.M.H. de Rooij
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Katerina Rohlenova
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Jermaine Goveia
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Luc Schoonjans
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong, P.R. China
- Mieke Dewerchin
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Guy Eelen
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium
- Xuri Li
- State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong, P.R. China
- Joanna Kalucka
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium; Department of Biomedicine, Aarhus University, Aarhus 8000, Denmark; Corresponding author
- Peter Carmeliet
- Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong, P.R. China; Department of Biomedicine, Aarhus University, Aarhus 8000, Denmark; Corresponding author
- Journal volume & issue
-
Vol. 2,
no. 3
p. 100523
Abstract
Summary: Endothelial cells (ECs) exhibit phenotypic and functional tissue specificities, critical for studies in the vascular field and beyond. Thus, tissue-specific methods for isolation of highly purified ECs are necessary. Kidney, spleen, and testis ECs are relevant players in health and diseases such as chronic kidney disease, acute kidney injury, myelofibrosis, and cancer. Here, we provide tailored protocols for rapid and reproducible EC purification established for scRNA sequencing from these adult murine tissues using the combination of magnetic- and fluorescence-activated cell sorting.For complete details on the use and execution of these protocols, please refer to Kalucka et al. (2020) and Dumas et al. (2020).
