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Sizing Up Extracellular DNA: Instant Chromatin Discharge From Cells When Placed in Serum-Free Conditions
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Biomedical and Clinical Sciences, Division of Clinical Chemistry.
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Health, Medicine and Caring Sciences, Division of Diagnostics and Specialist Medicine.ORCID iD: 0000-0002-3328-5060
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Biomedical and Clinical Sciences, Division of Cell Biology.ORCID iD: 0000-0001-5082-6423
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Biomedical and Clinical Sciences, Division of Cell Biology.
2020 (English)In: Frontiers in Cell and Developmental Biology, E-ISSN 2296-634X, Vol. 8, article id 634Article in journal (Refereed) Published
Abstract [en]

How do you wash cells? Three out of four of our colleagues use experimental procedures during everyday lab-bench work that can severely impair data interpretation depending on how cells are handled. We show here that a subpopulation (2–3%) of human leukocytes immediately induce a yet unclassified lytic cell death, concomitant with discharge of chromatin entities and cell elimination, when placed in protein-free solutions (i.e., PBS and HBSS). DNA release was not restricted to hematopoietic cells but occurred also in HEK293T cells. Albumin, fetal bovine serum, polyethylene glycol, and Pluronic F-68 supplements prevented chromatin discharge. Expelled chromatin was devoid of surrounding membranes but maintained its original nuclear shape, although ∼10 times enlarged. These structures differed from DNA appearance after osmotic or detergent-induced cell lysis. Besides sounding a cautionary note to the neutrophil extracellular trap (NET) research community, in which ∼50% of all published studies used protein-free media for NET-formation, our study also provides a rapid tool for analysis of chromatin organization.

Place, publisher, year, edition, pages
2020. Vol. 8, article id 634
Keywords [en]
cell death, chromatin structure, extracellular DNA, leukocytes, serum-free conditions
National Category
Cell Biology
Identifiers
URN: urn:nbn:se:liu:diva-169025DOI: 10.3389/fcell.2020.00634ISI: 000559279100001PubMedID: 32793591Scopus ID: 2-s2.0-85089086280OAI: oai:DiVA.org:liu-169025DiVA, id: diva2:1464373
Conference
Sizing Up Extracellular DNA: Instant Chromatin Discharge From Cells When Placed in Serum-Free ConditionsGiannis Spyrou, Daniel Appelgren, Anders Rosén, Björn IngelssonFront Cell Dev Biol. 2020; 8: 634. Published online 2020 Jul 22. doi: 10.3389/fcell.2020.00634PMCID: PMC7387414
Available from: 2020-09-05 Created: 2020-09-05 Last updated: 2021-05-06Bibliographically approved

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Spyrou, GiannisAppelgren, DanielRosén, AndersIngelsson, Björn

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