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open-access-imgOpen AccessThe expression of endogenous voltage‐gated potassium channels in HEK293 cells is affected by culture conditions
Author(s)
Ponce Arturo,
Castillo Aida,
Hinojosa Lorena,
MartinezRendon Jacqueline,
Cereijido Marcelino
Publication year2018
Publication title
physiological reports
Resource typeJournals
PublisherWiley-Blackwell
Abstract HEK 293 cells are widely used as a host for expression of heterologous proteins; yet, little care has been taken to characterize their endogenous membrane components, including ion channels. In this work, we aimed to describe the biophysical and pharmacological properties of endogenous, voltage‐dependent potassium currents ( IK v). We also examined how its expression depends on culture conditions. We used the electrophysiological technique of whole‐cell patch clamp to record ion currents from HEK 293 cells. We found that HEK cells express endogenous, voltage‐dependent potassium currents. We also found that diverse culture conditions, such as the passage number, the cell density, the type of serum that complements the culture media and the substratum, affect the magnitude and shape of IK v, resulting from the relative contribution of fast, slow, and noninactivating component currents. Incubation of cells in mature monolayers with trypsin– EDTA , notoriously reduces the magnitude and modifies the shape of voltage‐dependent potassium endogenous currents; nonetheless HEK cells recover IK v′s magnitude and shape within 6 h after replating, with a process that requires synthesis of new mRNA and protein subunits, as evidenced by the fact that actinomycin D and cycloheximide, inhibitors of synthesis of mRNA and protein, respectively, impair the recovery of IK v after trypsinization. In addition to be useful as a model expression system, HEK 293 may be useful to understand how cells regulate the density of ion channels on the membrane.
Subject(s)biochemistry , biology , cell culture , chemistry , computer science , endocrinology , endogeny , expression (computer science) , genetics , hek 293 cells , ion channel , microbiology and biotechnology , organic chemistry , potassium , potassium channel , programming language , receptor , voltage gated ion channel , voltage gated potassium channel
Keyword(s) 4AP , HEK293 , potassium channels , TEA , trypsin
Language(s)English
SCImago Journal Rank0.918
H-Index39
ISSN2051-817X
DOI10.14814/phy2.13663

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