Plasma Membrane Wounding and Repair Assays for Eukaryotic Cells
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Plasma Membrane Wounding and Repair Assays for Eukaryotic Cells. / Sønder, Stine Lauritzen; Ebstrup, Malene Laage; Dias, Catarina; Busk Heitmann, Anne Sofie; Nylandsted, Jesper.
I: Bio-protocol, Bind 12, Nr. 11, e4437, 2022.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Plasma Membrane Wounding and Repair Assays for Eukaryotic Cells
AU - Sønder, Stine Lauritzen
AU - Ebstrup, Malene Laage
AU - Dias, Catarina
AU - Busk Heitmann, Anne Sofie
AU - Nylandsted, Jesper
N1 - Publisher Copyright: © 2022 Bio-protocol LLC. All Rights Reserved.
PY - 2022
Y1 - 2022
N2 - Damage to the plasma membrane and loss of membrane integrity are detrimental to eukaryotic cells. It is, therefore, essential that cells possess an efficient membrane repair system to survive. However, the different cellular and molecular mechanisms behind plasma membrane repair have not been fully elucidated. Here, we present three complementary methods for plasma membrane wounding, and measurement of membrane repair and integrity. The first protocol is based on real time imaging of cell membrane repair kinetics in response to laser-induced injury. The second and third protocols are end point assays that provide a population-based measure of membrane integrity, after either mechanical injury by vortex mixing with glass beads, or by detergent-induced injury by digitonin in sublytic concentrations. The protocols can be applied to most adherent eukaryotic cells in culture, as well as cells in suspension.
AB - Damage to the plasma membrane and loss of membrane integrity are detrimental to eukaryotic cells. It is, therefore, essential that cells possess an efficient membrane repair system to survive. However, the different cellular and molecular mechanisms behind plasma membrane repair have not been fully elucidated. Here, we present three complementary methods for plasma membrane wounding, and measurement of membrane repair and integrity. The first protocol is based on real time imaging of cell membrane repair kinetics in response to laser-induced injury. The second and third protocols are end point assays that provide a population-based measure of membrane integrity, after either mechanical injury by vortex mixing with glass beads, or by detergent-induced injury by digitonin in sublytic concentrations. The protocols can be applied to most adherent eukaryotic cells in culture, as well as cells in suspension.
KW - Cell injury
KW - Imaging
KW - Membrane integrity
KW - Membrane wounding
KW - Plasma membrane damage
KW - Plasma membrane repair assays
U2 - 10.21769/BioProtoc.4437
DO - 10.21769/BioProtoc.4437
M3 - Journal article
C2 - 35799909
AN - SCOPUS:85133326001
VL - 12
JO - Bio-protocol
JF - Bio-protocol
SN - 2331-8325
IS - 11
M1 - e4437
ER -
ID: 316679741