Publikationsdatenbank

Protocol to dissociate healthy and infected murine- and hamster-derived lung tissue for single-cell transcriptome analysis (2023)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Pennitz, Peter

Goekeri, Cengiz

Trimpert, Jakob (WE 5)

Wyler, Emanuel

Ebenig, Aileen

Weissfuss, Chantal

Mühlebach, Michael D.

Witzenrath, Martin

Nouailles, Geraldine

Quelle

STAR Protocols

Bandzählung: 4

Heftzählung: 1

Seiten: Artikel 101957

ISSN: 2666-1667

Sprache

Englisch

Verweise

URL (Volltext): https://www.sciencedirect.com/science/article/pii/S2666166722008371

DOI: 10.1016/j.xpro.2022.101957

Pubmed: 36542521

Kontakt

Institut für Virologie

Robert-von-Ostertag-Str. 7-13
14163 Berlin
+49 30 838 51833
virologie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

In infectious disease research, single-cell RNA sequencing allows dissection of host-pathogen interactions. As a prerequisite, we provide a protocol to transform solid and complex organs such as lungs into representative diverse, viable single-cell suspensions. Our protocol describes performance of vascular perfusion, pneumonectomy, enzymatic digestion, and mechanical dissociation of lung tissue, as well as red blood cell lysis and counting of isolated cells. A challenge remains, however, to further increase the proportion of pulmonary endothelial cells without compromising on viability.