von Ashesh Chakraborty ; Marie Zöller ; Aydan Sardogan ; Markus Klotz ; Michal Mastalerz ; Hannah Marchi ; Raphael Meixner ; Rudolf A. Hatz ; Jürgen Behr ; Anne Hilgendorff ; Misako Nakayama ; Claudia Staab-Weijnitz
von Steven H. Abman ; Sylvia M. Nikkho ; Rolf M. F. Berger ; Maria Jesus del Cerro ; Eric D. Austin ; Maurice Beghetti ; Dunbar Ivy ; Megan Griffiths ; Anne Hilgendorff ; Steven H. Kawut ; Usha S. Krishnan ; Mary P. Mullen ; Shahin Moledina ; Bernard Thébaud ; Norman Stockbridge
von Saahithi Mallapragada ; Ruqian Lyu ; Arianna L. Williams-Katek ; Brandon K. Fischer ; Annika Vannan ; Niran Hadad ; Evan D. Mee ; Shawyon P. Shirazi ; Christopher S. Jetter ; Nicholas M. Negretti ; Anne Hilgendorff ; Laurie C. Eldredge ; Gail H. Deutsch ; Davis J. McCarthy ; Jonathan A. Kropski ; Jennifer M. S. Sucre ; Nicholas E. Banovich
von Francesco Palumbo ; Mis̆a Gunjak ; Patty J. Lee ; Stefan Günther ; Anne Hilgendorff ; István Vadász ; Susanne Herold ; Werner Seeger ; Christian Axel Mühlfeld ; Rory E. Morty
Flow cytometry and fluorescence-activated cell sorting are widely used to study endothelial cells, for which the generation of viable single-cell suspensions is an essential first step. Two enzymatic approaches, collagenase A and dispase, are widely employed for endothelial cell isolation. In this study, the utility of both enzymatic approaches, alone and in combination, for endothelial cell isolation from juvenile and adult mouse lungs was assessed, considering the number, viability, and subtype composition of recovered endothelial cell pools. Collagenase A yielded an 8-12-fold superior recovery of viable endothelial cells from lung tissue from developing mouse pups, compared to dispase, although dispase proved superior in efficiency for epithelial cell recovery. Single-cell RNA-Seq revealed that the collagenase A approach yielded a diverse endothelial cell subtype composition of recovered endothelial cell pools, with broad representation of arterial, capillary, venous, and lymphatic lung endothelial cells; while the dispase approach yielded a recovered endothelial cell pool highly enriched for one subset of general capillary endothelial cells, but poor representation of other endothelial cells subtypes. These data indicate that tissue dissociation markedly influences the recovery of endothelial cells, and the endothelial subtype composition of recovered endothelial cell pools, as assessed by single-cell RNA-Seq.
von Octavia-Andreea Ciora ; Tanja Seegmüller ; Johannes S. Fischer ; Theresa Wirth ; Friederike Häfner ; Sophia Stoecklein ; Andreas W. Flemmer ; Kai Förster ; Alida Kindt ; Dirk Bassler ; Christian F. Poets ; Narges Ahmidi ; Anne Hilgendorff
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