STAR Protocols (Sep 2021)
Protocols for endothelial cell isolation from mouse tissues: brain, choroid, lung, and muscle
- Nadine V. Conchinha,
- Liliana Sokol,
- Laure-Anne Teuwen,
- Koen Veys,
- Sébastien J. Dumas,
- Elda Meta,
- Melissa García-Caballero,
- Vincent Geldhof,
- Rongyuan Chen,
- Lucas Treps,
- Mila Borri,
- 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
- 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
- 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 and 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
- 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
- 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
- Rongyuan Chen
- State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong, P.R. China
- 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
- 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
- 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. 100508
Abstract
Summary: Endothelial cells (ECs) harbor distinct phenotypical and functional characteristics depending on their tissue localization and contribute to brain, eye, lung, and muscle diseases such as dementia, macular degeneration, pulmonary hypertension, and sarcopenia. To study their function, isolation of pure ECs in high quantities is crucial. Here, we describe protocols for rapid and reproducible blood vessel EC purification established for scRNA sequencing from murine tissues using mechanical and enzymatic digestion followed by magnetic and fluorescence-activated cell sorting.For complete details on the use and execution of these protocol, please refer to Kalucka et al. (2020), Rohlenova et al. (2020), and Goveia et al. (2020).