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Changes in morphology and inward rectifier currents in human atrial myocytes depend on culture conditions
Authors:Herbert M. Himmel  Martina Pietsch  Uwe Streller  Eva M. Graf  Ursula Ravens
Affiliation:Bayer AG, Abt Sicherheitsstudien, Wuppertal, Germany.
Abstract:
Human atrial myocytes were cultured under systematically varied conditions in order to obtain stable cells for future gene manipulation. Transient (Ito) and sustained outward current (Iso), and voltage- and muscarinic receptor-activated inward rectifier K+ currents (IK1, IK,ACh) were measured in freshly isolated cells and after 5 days in culture. Myocytes were grown on polylysin or laminin in medium with or without 10 % serum (medium+S, medium-S). Cultured myocytes dedifferentiated to a greater extent in medium+S than medium-S, but independent of the chemical nature of the adherence surface. Apparent surface area increased in medium+S, whereas membrane capacitance declined under all culture conditions. Ito of myocytes cultured in medium-S was increased. Myocytes grown on polylysin and laminin exhibited reduced IK1 current density. Under all culture conditions, IK,ACh was attenuated with carbachol but hardly affected with sphingosine-1-phosphate as agonists. In conclusion, morphological and electrophysiological changes depended on serum in the culture medium rather than on adherence surface being coated with laminin or polylysin. Received: 4 October 2001, Returned for revision: 25 October 2001, Revision received: 4 July 2002, Accepted: 25 July 2002 H. M. H. and M. P. contributed equally to this work. Correspondence to: U. Ravens
Keywords:K+ currents –   sphingosine-1-phosphate –   serum-free culture –   polylysin –   laminin
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