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Verfasst von:Babin-Ebell Gonçalves, Anna [VerfasserIn]   i
 Mao, Yifang [VerfasserIn]   i
 Baljkas, Tinja [VerfasserIn]   i
 Wiedmann, Felix Tobias [VerfasserIn]   i
 Eis, Larissa [VerfasserIn]   i
 Pilz, Franziska [VerfasserIn]   i
 Winkler, Manuel [VerfasserIn]   i
 Kürschner, Sina Wietje [VerfasserIn]   i
 Hoffarth, Marlene [VerfasserIn]   i
 Funaya, Charlotta [VerfasserIn]   i
 Shahidi, Réza [VerfasserIn]   i
 Géraud, Cyrill [VerfasserIn]   i
 Wu, Chi-Chung [VerfasserIn]   i
 Schmidt, Constanze [VerfasserIn]   i
 Goerdt, Sergij [VerfasserIn]   i
 Reiners-Koch, Philipp-Sebastian [VerfasserIn]   i
 Singhal, Mahak [VerfasserIn]   i
Titel:A perfusion-independent high-throughput method to isolate liver sinusoidal endothelial cells
Verf.angabe:Anna Babin-Ebell Gonçalves, Yifang Mao, Tinja Baljkas, Felix Wiedmann, Larissa Eis, Franziska Pilz, Manuel Winkler, Sina W. Kürschner-Zacharias, Marlene Hoffarth, Charlotta Funaya, Réza Shahidi, Cyrill Géraud, Chi-Chung Wu, Constanze Schmidt, Sergij Goerdt, Philipp-Sebastian Reiners-Koch & Mahak Singhal
E-Jahr:2025
Jahr:8. Januar 2025
Umfang:10 S.
Fussnoten:Gesehen am 27.03.2025
Titel Quelle:Enthalten in: Communications biology
Ort Quelle:London : Springer Nature, 2018
Jahr Quelle:2025
Band/Heft Quelle:8(2025), 1, Artikel-ID 22, Seite 1-10
ISSN Quelle:2399-3642
Abstract:Liver sinusoidal endothelial cells (LSECs) critically regulate homeostatic liver function and liver pathogenesis. However, the isolation of LSECs remains a major technological bottleneck in studying molecular mechanisms governing LSEC functions. Current techniques to isolate LSECs, relying on perfusion-dependent liver digestion, are cumbersome with limited throughput. We here describe a perfusion-independent high-throughput procedure to isolate LSECs with high purity. Indifferently from previous perfusion-independent approaches, chopped liver tissue was incubated in the digestion mix for 30 minutes with intermittent mixing with a serological pipette. This led to the safeguarding of LSEC integrity and yielded 10 ± 1.0 million LSECs per adult mouse liver, which is far higher than previous perfusion-independent protocols and comparable yield to established perfusion-dependent protocols for isolating LSECs. Combining magnetic and fluorescence-activated cell sorting (FACS), LSECs from different zones of the hepatic sinusoid can now be isolated in high numbers in less than two hours for downstream applications including proteomics. Our protocol enables the isolation of LSECs from fibrotic liver tissues from mice and healthy liver tissues from higher vertebrate species (pigs), where traditional perfusion-based digestion protocols have very limited application. In conclusion, these technical advancements reduce post-mortem changes in the LSEC state and aid in reliable investigation of LSEC functions.
DOI:doi:10.1038/s42003-025-07458-5
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

kostenfrei: Volltext: https://doi.org/10.1038/s42003-025-07458-5
 kostenfrei: Volltext: https://www.nature.com/articles/s42003-025-07458-5
 DOI: https://doi.org/10.1038/s42003-025-07458-5
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:Angiogenesis
 Liver
K10plus-PPN:1920665404
Verknüpfungen:→ Zeitschrift

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