caa a22 4500 445822848 CHVBK 20180317145249.0 cr unu---uuuuu 170323e20110101xx s 000 0 eng 10.1007/s10616-010-9332-7 doi (NATIONALLICENCE)springer-10.1007/s10616-010-9332-7 Rational method in the repetitive calcium oscillation measurement in wild type human epithelial kidney cells [Elektronische Daten] [Xian Jing, Lihong Chen, Shuang Ren, Dali Luo] Cells stimulated with physiological stimuli usually exhibit oscillations in cytosolic Ca2+ concentration ([Ca2+]i), a signal playing central roles in regulation of various cellular processes. For explicating their unknown mechanisms, studies are commonly conducted in single cells from several cell lines, in particular the human epithelial kidney (HEK293) cell line. However, [Ca2+]i oscillating responses to agonists in vitro are found difficult to be induced and varied with different types of cells and agonists. This study shows that treatment of the wild type HEK293 cells with low concentrations of carbachol (1-10μM), an agonist of the muscarinic receptor, resulted in non-oscillated but sustained [Ca2+]i increase by loading the cells with 1 μM fura2/AM. However, repetitive and long lasting [Ca2+]i oscillations could be induced in 31.1% of the tested cells loaded with 0.1μM fura2/AM. Additionally, the occurrence of the typical Ca2+ spikes further increased to 47.2% and 60.7% when the Ca2+ concentration in the bathing medium was decreased from 1.8 mM to 1.5mM and the medium temperature was set to 35±1°C from 22±2°C. Therefore, this study provides a useful approach for measuring [Ca2+]i oscillatory response to relevant physiological stimulation in a wild type cell line through the adjustments of the concentrations adopted for the Ca2+ indicator and extracellular medium Ca2+ and of the temperature set for the experiment. Springer Science+Business Media B.V., 2011 Ca2+ oscillations nationallicence Fura2/AM nationallicence Human epithelial kidney cells nationallicence Carbachol nationallicence Jing Xian Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut Chen Lihong Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut Ren Shuang Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut Luo Dali Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut Cytotechnology Springer Netherlands 63/1(2011-01-01), 81-88 0920-9069 63:1<81 2011 63 10616 https://doi.org/10.1007/s10616-010-9332-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-010-9332-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Jing Xian Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Chen Lihong Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Ren Shuang Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Luo Dali Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, 100069, Beijing, People's Republic of China aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/1(2011-01-01), 81-88 0920-9069 63:1<81 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer