Open AccessMembrane permeability changes induce hyperpolarization in transformed lymphoid cells under high‐density culture conditions
Author(s) -
Márián Teréz,
Balkay László,
Krasznai Zoltán,
Trón Lajos
Publication year - 2000
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/1097-0320(20001101)41:3<186::aid-cyto5>3.0.co;2-j
Subject(s) - intracellular , membrane potential , cell culture , permeability (electromagnetism) , hyperpolarization (physics) , membrane , membrane permeability , biophysics , flow cytometry , chemistry , potassium , cell , sodium , cell membrane , microbiology and biotechnology , biology , biochemistry , stereochemistry , organic chemistry , nuclear magnetic resonance spectroscopy , genetics
Background Membrane potential changes in cells from the human lymphoid B cell line, JY, evoked by increasing cell density in culture were investigated, as data published on other cell types are controversial. An attempt was also made to clear the underlying mechanism. Methods Nonadherent JY cells were isolated from high‐density plateau‐phase cultures (type A cells), medium‐density log‐phase cultures (type B cells), and low‐density lag‐phase cultures (type C cells). They were analyzed for transmembrane potential, intracellular free concentration of potassium and sodium, membrane permeability for monovalent cations, cell cycle distribution by measuring DNA content, and glucose uptake. Results C type cells proved to be relatively depolarized (−41 ± 3 mV) and cells obtained from the highest density cultures hyperpolarized (−60 ± 3 mV). Intracellular concentrations ([ K ] i = 92–97 mM and [Na] i = 34–35 mM ) were almost identical for each type of cell. The sodium/potassium permeability constant ratio in the A and C type of cells was 0.047 and 0.094, respectively. High‐density culture conditions resulted in a pronounced G 1 ‐phase arrest. Conclusions Differences in the membrane potential values induced by high‐density culture conditions were maintained by changes in the membrane permeability for the monovalent cations. Cytometry 41:186‐192,2000. © 2000 Wiley‐Liss, Inc.