title: Electrical stimulation of human embryonic stem cells: cardiac differentiation and the generation of reactive oxygen species.
abstract: Exogenous electric fields have been implied in cardiac differentiation of mouse embryonic stem cells and the generation of reactive oxygen species (ROS). In this work, we explored the effects of electrical field stimulation on ROS generation and cardiogenesis in embryoid bodies (EBs) derived from human embryonic stem cells (hESC, line H13), using a custom-built electrical stimulation bioreactor. Electrical properties of the bioreactor system were characterized by electrochemical impedance spectroscopy (EIS) and analysis of electrical currents. The effects of the electrode material (stainless steel, titanium-nitride-coated titanium, titanium), length of stimulus (1 and 90 s) and age of EBs at the onset of electrical stimulation (4 and 8 days) were investigated with respect to ROS generation. The amplitude of the applied electrical field was 1 V/mm. The highest rate of ROS generation was observed for stainless steel electrodes, for signal duration of 90 s and for 4-day-old EBs. Notably, comparable ROS generation was achieved by incubation of EBs with 1 nM H(2)O(2). Cardiac differentiation in these EBs was evidenced by spontaneous contractions, expression of troponin T and its sarcomeric organization. These results imply that electrical stimulation plays a role in cardiac differentiation of hESCs, through mechanisms associated with the intracellular generation of ROS.
authors.affiliation: Department of Biomedical Engineering, Columbia University, New York, NY, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Serena
authors.forename: Elisa
authors.initials: E
authors.lastname: Figallo
authors.forename: Nina
authors.initials: N
authors.lastname: Tandon
authors.forename: Christopher
authors.initials: C
authors.lastname: Cannizzaro
authors.forename: Sharon
authors.initials: S
authors.lastname: Gerecht
authors.forename: Nicola
authors.initials: N
authors.lastname: Elvassore
authors.forename: Gordana
authors.initials: G
authors.lastname: Vunjak-Novakovic
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002470:Cell Survival; D004558:Electric Stimulation; D004560:Electricity; D004566:Electrodes; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012518:Sarcomeres / Q000378:metabolism; D013193:Stainless Steel / Q000737:chemistry; D014025:Titanium / Q000737:chemistry; D020107:Troponin T / Q000378:metabolism
chemical_list: D005452:Fluoresceins; D017382:Reactive Oxygen Species; D020107:Troponin T; D013193:Stainless Steel; C029569:diacetyldichlorofluorescein; C041500:titanium nitride; D006861:Hydrogen Peroxide; D014025:Titanium