Sizing Up Extracellular DNA: Instant Chromatin Discharge From Cells When Placed in Serum-Free Conditions
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
2020-09-052020-09-052021-05-06Bibliographically approved