Isolation of Human Neutrophils

All experiments with human neutrophils were approved by the Ethics Committee of the University Medical Center (UMG) Göttingen (protocol number: 29/1/17). Neutrophils were isolated from fresh venous blood of healthy donors. Beforehand, all donors were fully informed about possible risks, and their informed consent was obtained in writing. The consent could be withdrawn at any time during the study. Blood was collected in S-Monovettes EDTA (7.5 ml, Sarstedt), and neutrophils were isolated according to previously published protocols based on histopaque 1119 (Sigma Aldrich) as well as Percoll (GE Healthcare) density gradients (17 (link), 42 (link)). Neutrophils were resuspended in HBSS^−Ca2+/Mg2+ and further diluted in the desired medium as described in the appropriate methods sections and figure legends. Purity of the cell preparation was >95% as assessed by cytospin (Cytospin 2 centrifuge, Shanson) and Diff Quick staining (Medion, Diagnostics).

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Neubert E., Bach K.M., Busse J., Bogeski I., Schön M.P., Kruss S, & Erpenbeck L. (2019). Blue and Long-Wave Ultraviolet Light Induce in vitro Neutrophil Extracellular Trap (NET) Formation. Frontiers in Immunology, 10, 2428.

Publication 2019

S-Monovettes EDTA histopaque 1119 Percoll

Blood Cell Diagnostics Donors Donors blood Edta Hbss Histopaque Human Neutrophils Percoll Venous

Corresponding Organization :

Other organizations : University of Göttingen, Nanoscale Microscopy and Molecular Physiology of the Brain Cluster of Excellence 171 — DFG Research Center 103

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Variable analysis

independent variables

Isolation of neutrophils from fresh venous blood of healthy donors

dependent variables

Purity of the cell preparation

control variables

Obtaining informed consent from donors
Collecting blood in S-Monovettes EDTA (7.5 ml, Sarstedt)
Isolating neutrophils according to previously published protocols based on histopaque 1119 (Sigma Aldrich) and Percoll (GE Healthcare) density gradients
Resuspending neutrophils in HBSS-Ca2+/Mg2+ and further diluting in the desired medium

controls

Positive control: Not specified
Negative control: Not specified

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