- Email: [email protected]
Pulsed electromagnetic fields have neuroprotective effects on cultured dopaminergic neurons
ABSTRACTS / Experimental Neurology 198 (2006) 558 – 597
Pulsed electromagnetic fields have neuroprotective effects on cultured dopaminergic neurons D. Casper 1, E. Taub 1, L. Alammar 1, A. Pidel 1, A.A. Pilla 2 1 Neurosurgery Lab, Montefiore Medical Center and the Albert Einstein College of Medicine, Bronx, USA 2 Department of Biomedical Engineering, Columbia University, New York, USA Parkinson’s disease is characterized by the loss of dopaminergic neurons. Various strategies to attenuate neuronal loss may be successful under experimental conditions, but their usefulness is ultimately limited by problems in delivering neuroprotective agents to the brain, undesirable side effects, and the chronic nature of the disease itself. This study was undertaken to explore the potential for pulsed electromagnetic fields (PEMF) to increase the viability of dopaminergic neurons. Using primary cultures of rat embryonic midbrain, we employed pulsed radiofrequency signals and tested whether they could increase survival in response to quisqualic acid, a glutamate receptor agonist, 6-hydroxydopamine, a dopaminergic neurotoxin, or serum deprivation. Cultures were exposed to PEMF signals (1 – 10 ms bursts of a 27.12 MHz sinusoidal wave at 1– 5 bursts/sec at 0.05 Gauss amplitude) following acute and chronic regimens. At 6 days in vitro, cells were treated with toxins. After 1– 2 days, cultures were fixed for tyrosine hydroxylase immunocytochemistry to identify dopaminergic neurons or harvested for determination of cyclic GMP content. Conditioned media were collected to determine nitric oxide (NO) content. PEMF increased the survival of dopaminergic neurons in all three neurotoxicity paradigms. These effects were sensitive to changes in signal burst widths and repetition rates. In addition, cyclic GMP levels increased 4-fold. Others have shown that PEMF signals can affect the binding of calcium ions to calmodulin, inducing NO production, which up-regulates the formation of cyclic GMP. Although NO levels remained constant, a transient role for NO was supported by using l-NAME, an NO synthesis inhibitor, which attenuated PEMF-induced cyclic GMP formation. Taken together, results suggest that PEMFs can protect dopaminergic neurons from several types of toxicity. Further experiments with inhibitors of this pathway will determine more definitively whether it mediates the neuroprotection. (This work was supported by NIH grant R21 NS052576-01).
563
4
Department of Anatomy, USF College of Medicine, Tampa, Florida 5 USF College of Medicine and Saneron CCEL Therapeutics, Tampa, Florida 6 Department of Pharmacology and Therapeutics, USF College of Medicine, Tampa, Florida 7 Department of Pathology, USF College of Medicine, Tampa, Florida 9 Department of Physiology and Biophysics, USF College of Medicine, Tampa, Florida Human umbilical cord blood (HUCB) cells are capable of inducing recovery in numerous models of central nervous system (CNS) disease and injury such as spinal cord injury, stroke, amyotrophic lateral sclerosis, and other age-associated diseases. The aim of this study was to examine the trophic effects of HUCB cells on mature hippocampal neurons in vitro. We co-cultured the mononuclear fraction of HUCB cells (HUCBmnf) with hippocampus neural cells derived from young adult rat brain (7 months of age). Prior to placing the HUCBmnf cells in culture with the hippocampal neurons, HUCBmnf were cultured with DMEM and 20% FBS, with or without pretreatment with human epithelial growth factor (hEGF), human basic fibrous growth factor (hFGFb) and human stem cell factors (hSCF). Hippocampal neurons and HUCBmnf were then cultured under conditions appropriate for neuronal growth for 14, 21, 28 and 35 days in vitro (DIV). Immunochemistry staining was employed to identify neurons (MAP2+) and glial cells (GFAP+) as well as arborization of neurites. The average number of MAP2+ hippocampal neurons cells in the co-cultures was significantly higher than in the control group (hippocampal mono-cultures). These MAP2+ hippocampal cells in co-culture were richly arborized, especially after 21 and 28 DIV. At the same time, we also noticed that HUCBmnf cells have an anti-senescence effect on the hippocampal cultures, with 90% of the hippocampal cells in co-culture surviving for up to 35 days. However, number of hippocampus cells in the control group declined over time in mono-culture especially at the later time points when large numbers of cells were dying. Ongoing studies will further explore the mechanism underlying this trophic effect of HUCBmnf cells on the growth of hippocampal neurons. (Supported by a NIH grant R01AG020927 to AEW. SGD, CDS, PRS, and AEW are consultants to Saneron CCEL Therapeutics).
doi:10.1016/j.expneurol.2006.02.028
doi:10.1016/j.expneurol.2006.02.029
Effect of human umbilical cord blood cells on adult hippocampal neurons in vitro N. Chen 1,2, A Simmens 1,2, J. Newcomb 1,2, S. Kamath 3, S. Garbuzova-Davis 1,2,7,9, C.D. Sanberg 5, P.R. Sanberg 1,2,5,9, P.C. Bickford 1,2,6, A.E. Willing 1,2,4,6,7 1 Center of Excellence for Aging and Brain Repair, USF College of Medicine, Tampa, Florida 2 Department of Neurosurgery, USF College of Medicine, Tampa, Florida 3 Department of Neurology, USF College of Medicine, Tampa, Florida
Isolation, culture and differentiation of neural stem cells from hippocampus of mouse embryo W.Y. Chen 1,2, F.G. Meng 1,2 1 Department of Neurosurgery, Tianjin Huanhu Hospital, Tianjin, China 2 Tianjin Neurosurgical Institute2, Tianjin, China Objective: To establish the isolation, culture and differentiation of neural stem cells from hippocampus of mouse embryo in vitro. Methods: Mouse embryonic hippocampus (E13) was isolated and cultured in DMEM/F-12 with growth factor EGF and
Pulsed electromagnetic fields have neuroprotective effects on cultured dopaminergic neurons D. Casper 1, E. Taub 1, L. Alammar 1, A. Pidel 1, A.A. Pilla 2 1 Neurosurgery Lab, Montefiore Medical Center and the Albert Einstein College of Medicine, Bronx, USA 2 Department of Biomedical Engineering, Columbia University, New York, USA Parkinson’s disease is characterized by the loss of dopaminergic neurons. Various strategies to attenuate neuronal loss may be successful under experimental conditions, but their usefulness is ultimately limited by problems in delivering neuroprotective agents to the brain, undesirable side effects, and the chronic nature of the disease itself. This study was undertaken to explore the potential for pulsed electromagnetic fields (PEMF) to increase the viability of dopaminergic neurons. Using primary cultures of rat embryonic midbrain, we employed pulsed radiofrequency signals and tested whether they could increase survival in response to quisqualic acid, a glutamate receptor agonist, 6-hydroxydopamine, a dopaminergic neurotoxin, or serum deprivation. Cultures were exposed to PEMF signals (1 – 10 ms bursts of a 27.12 MHz sinusoidal wave at 1– 5 bursts/sec at 0.05 Gauss amplitude) following acute and chronic regimens. At 6 days in vitro, cells were treated with toxins. After 1– 2 days, cultures were fixed for tyrosine hydroxylase immunocytochemistry to identify dopaminergic neurons or harvested for determination of cyclic GMP content. Conditioned media were collected to determine nitric oxide (NO) content. PEMF increased the survival of dopaminergic neurons in all three neurotoxicity paradigms. These effects were sensitive to changes in signal burst widths and repetition rates. In addition, cyclic GMP levels increased 4-fold. Others have shown that PEMF signals can affect the binding of calcium ions to calmodulin, inducing NO production, which up-regulates the formation of cyclic GMP. Although NO levels remained constant, a transient role for NO was supported by using l-NAME, an NO synthesis inhibitor, which attenuated PEMF-induced cyclic GMP formation. Taken together, results suggest that PEMFs can protect dopaminergic neurons from several types of toxicity. Further experiments with inhibitors of this pathway will determine more definitively whether it mediates the neuroprotection. (This work was supported by NIH grant R21 NS052576-01).
563
4
Department of Anatomy, USF College of Medicine, Tampa, Florida 5 USF College of Medicine and Saneron CCEL Therapeutics, Tampa, Florida 6 Department of Pharmacology and Therapeutics, USF College of Medicine, Tampa, Florida 7 Department of Pathology, USF College of Medicine, Tampa, Florida 9 Department of Physiology and Biophysics, USF College of Medicine, Tampa, Florida Human umbilical cord blood (HUCB) cells are capable of inducing recovery in numerous models of central nervous system (CNS) disease and injury such as spinal cord injury, stroke, amyotrophic lateral sclerosis, and other age-associated diseases. The aim of this study was to examine the trophic effects of HUCB cells on mature hippocampal neurons in vitro. We co-cultured the mononuclear fraction of HUCB cells (HUCBmnf) with hippocampus neural cells derived from young adult rat brain (7 months of age). Prior to placing the HUCBmnf cells in culture with the hippocampal neurons, HUCBmnf were cultured with DMEM and 20% FBS, with or without pretreatment with human epithelial growth factor (hEGF), human basic fibrous growth factor (hFGFb) and human stem cell factors (hSCF). Hippocampal neurons and HUCBmnf were then cultured under conditions appropriate for neuronal growth for 14, 21, 28 and 35 days in vitro (DIV). Immunochemistry staining was employed to identify neurons (MAP2+) and glial cells (GFAP+) as well as arborization of neurites. The average number of MAP2+ hippocampal neurons cells in the co-cultures was significantly higher than in the control group (hippocampal mono-cultures). These MAP2+ hippocampal cells in co-culture were richly arborized, especially after 21 and 28 DIV. At the same time, we also noticed that HUCBmnf cells have an anti-senescence effect on the hippocampal cultures, with 90% of the hippocampal cells in co-culture surviving for up to 35 days. However, number of hippocampus cells in the control group declined over time in mono-culture especially at the later time points when large numbers of cells were dying. Ongoing studies will further explore the mechanism underlying this trophic effect of HUCBmnf cells on the growth of hippocampal neurons. (Supported by a NIH grant R01AG020927 to AEW. SGD, CDS, PRS, and AEW are consultants to Saneron CCEL Therapeutics).
doi:10.1016/j.expneurol.2006.02.028
doi:10.1016/j.expneurol.2006.02.029
Effect of human umbilical cord blood cells on adult hippocampal neurons in vitro N. Chen 1,2, A Simmens 1,2, J. Newcomb 1,2, S. Kamath 3, S. Garbuzova-Davis 1,2,7,9, C.D. Sanberg 5, P.R. Sanberg 1,2,5,9, P.C. Bickford 1,2,6, A.E. Willing 1,2,4,6,7 1 Center of Excellence for Aging and Brain Repair, USF College of Medicine, Tampa, Florida 2 Department of Neurosurgery, USF College of Medicine, Tampa, Florida 3 Department of Neurology, USF College of Medicine, Tampa, Florida
Isolation, culture and differentiation of neural stem cells from hippocampus of mouse embryo W.Y. Chen 1,2, F.G. Meng 1,2 1 Department of Neurosurgery, Tianjin Huanhu Hospital, Tianjin, China 2 Tianjin Neurosurgical Institute2, Tianjin, China Objective: To establish the isolation, culture and differentiation of neural stem cells from hippocampus of mouse embryo in vitro. Methods: Mouse embryonic hippocampus (E13) was isolated and cultured in DMEM/F-12 with growth factor EGF and