Physiological And Molecular Genetic Effects Of Time Varying Electromagnetic Fields (TVEMF) On Human Neuronal Cells

Abstract

Many studies have been conducted to illuminate the potential positive benefits and/or negative effects of extremely low frequency (ELF) electromagnetic fields on human tissues. To date the results of these studies have yet to reveal the underlying cellular mechanisms for the demonstrated outcomes. PURPOSE This investigation details the development of model tissue systems and associated equipment for the proliferation two (2D) and three-dimensional (3D), in rotating wall vessels (RWVs), human neural progenitor cell tissue-like assemblies (TLAs) within a culture medium facilitated by a time-varying electromagnetic field (TVEMF) for the eventual purpose of In Vivo tissue regeneration. METHODS Normal human neural progenitor (NHNP) cells and culture medium are contained within a two or three-dimensional culture vessel, and the electromagnetic field is emitted from either an electrode or coil. Cells were exposed to a 10 Hz, 6 mA pulse width modulated square wave for periods of 17 up to 170 days. Duty cycles were in general 80%. RESULTS Data obtained from long duration stabilized studies realized increases in glucose utilization concomitant with cellular proliferation of 2.5–4 times the rate of the control cells. Appropriate cellular markers, beta tubulinn III, NCS-1 and others as determined by immunohistochemistry continued to be expressed throughout the experiments. At experiment termination as determined by flowcytometry, >83% of the treated population of cells were CD90 and CD133 positive, while 46% remained CXCR4 positive Affymetric gene chip analyses of 12,000 human genes demonstrated the ability to upregulate growth response genes and down regulate growth regulatory and suppressor genes. Extensive multicolor chromosome analysis by mFish demonstrated the treated population to be perfectly normal at 170 days. CONCLUSIONS As is clearly demonstrated in the human body, the bioelectric, biochemical process of electrical nerve stimulation is a documented reality. The present investigation demonstrates that a similar phenomenon can be potentiated in a synthetic atmosphere, i.e., 2D or 3D in rotating wall cell culture vessels. This electrical potentiation can be used for a number of purposes including developing tissues for transplantation, repair of traumatized tissues and as a modulator of some neurodegenerative diseases. These studies further provide methods to promote neural tissue regeneration while simultaneously studying and understanding the necessary fundamental mechanisms and elements of tissue regeneration.