A method to deliver patterned electrical impulses to Schwann cells cultured on an artificial axon
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摘要
Information from the brain travels back and forth along peripheral nerves in the form of electrical impulses generated by neurons and these impulses have repetitive patterns. Schwann cells in peripheral nerves receive molecular signals from axons to coordinate the process of myelination. There is evidence, however,that non-molecular signals play an important role in myelination in the form of patterned electrical impulses generated by neuronal activity. The role of patterned electrical impulses has been investigated in the literature using co-cultures of neurons and myelinating cells. The co-culturing method, however, prevents the uncoupling of the direct effect of patterned electrical impulses on myelinating cells from the indirect effect mediated by neurons. To uncouple these effects and focus on the direct response of Schwann cells,we developed an in vitro model where an electroconductive carbon fiber acts as an artificial axon. The fiber provides only the biophysical characteristics of an axon but does not contribute any molecular signaling.In our "suspended wire model", the carbon fiber is suspended in a liquid media supported by a 3D printed scaffold. Patterned electrical impulses are generated by an Arduino 101 microcontroller. In this study, we describe the technology needed to set-up and eventually replicate this model. We also report on our initial in vitro tests where we were able to document the adherence and ensheath of human Schwann cells to the carbon fiber in the presence of patterned electrical impulses(hSCs were purchased from ScienCell Research Laboratories, Carlsbad, CA, USA; ScienCell fulfills the ethic requirements, including donor's consent). This technology will likely make feasible to investigate the response of Schwann cells to patterned electrical impulses in the future.
Information from the brain travels back and forth along peripheral nerves in the form of electrical impulses generated by neurons and these impulses have repetitive patterns. Schwann cells in peripheral nerves receive molecular signals from axons to coordinate the process of myelination. There is evidence, however,that non-molecular signals play an important role in myelination in the form of patterned electrical impulses generated by neuronal activity. The role of patterned electrical impulses has been investigated in the literature using co-cultures of neurons and myelinating cells. The co-culturing method, however, prevents the uncoupling of the direct effect of patterned electrical impulses on myelinating cells from the indirect effect mediated by neurons. To uncouple these effects and focus on the direct response of Schwann cells,we developed an in vitro model where an electroconductive carbon fiber acts as an artificial axon. The fiber provides only the biophysical characteristics of an axon but does not contribute any molecular signaling.In our "suspended wire model", the carbon fiber is suspended in a liquid media supported by a 3D printed scaffold. Patterned electrical impulses are generated by an Arduino 101 microcontroller. In this study, we describe the technology needed to set-up and eventually replicate this model. We also report on our initial in vitro tests where we were able to document the adherence and ensheath of human Schwann cells to the carbon fiber in the presence of patterned electrical impulses(hSCs were purchased from ScienCell Research Laboratories, Carlsbad, CA, USA; ScienCell fulfills the ethic requirements, including donor's consent). This technology will likely make feasible to investigate the response of Schwann cells to patterned electrical impulses in the future.
Information from the brain travels back and forth along peripheral nerves in the form of electrical impulses generated by neurons and these impulses have repetitive patterns. Schwann cells in peripheral nerves receive molecular signals from axons to coordinate the process of myelination. There is evidence, however,that non-molecular signals play an important role in myelination in the form of patterned electrical impulses generated by neuronal activity. The role of patterned electrical impulses has been investigated in the literature using co-cultures of neurons and myelinating cells. The co-culturing method, however, prevents the uncoupling of the direct effect of patterned electrical impulses on myelinating cells from the indirect effect mediated by neurons. To uncouple these effects and focus on the direct response of Schwann cells,we developed an in vitro model where an electroconductive carbon fiber acts as an artificial axon. The fiber provides only the biophysical characteristics of an axon but does not contribute any molecular signaling.In our "suspended wire model", the carbon fiber is suspended in a liquid media supported by a 3D printed scaffold. Patterned electrical impulses are generated by an Arduino 101 microcontroller. In this study, we describe the technology needed to set-up and eventually replicate this model. We also report on our initial in vitro tests where we were able to document the adherence and ensheath of human Schwann cells to the carbon fiber in the presence of patterned electrical impulses(hSCs were purchased from ScienCell Research Laboratories, Carlsbad, CA, USA; ScienCell fulfills the ethic requirements, including donor's consent). This technology will likely make feasible to investigate the response of Schwann cells to patterned electrical impulses in the future.
引文
Althaus HH,Montz H,Neuhoff V,Schwartz P(1984)Isolation and cultivation of mature oligodendroglial cells.Naturwissenschaften 71:309-315.
Bechler ME,Byrne L,Ffrench-Constant C(2015)CNS Myelin Sheath Lengths Are an Intrinsic Property of Oligodendrocytes.Curr Biol25:2411-2416.
Bechler ME,Swire M,Ffrench-Constant C(2018)Intrinsic and adaptive myelination-A sequential mechanism for smart wiring in the brain.Dev Neurobiol 78:68-79.
Bullock PN,Rome LH(1990)Glass micro-fibers:a model system for study of early events in myelination.J Neurosci Res 27:383-393.
Buonanno A,Fields RD(1999)Gene regulation by patterned electrical activity during neural and skeletal muscle development.Curr Opin Neurobiol9:110-120.
Chang KJ,Redmond SA,Chan JR(2016)Remodeling myelination:implications for mechanisms of neural plasticity.Nat Neurosci 19:190-197.
Coggan JS,Bittner S,Stiefel KM,Meuth SG,Prescott SA(2015)Physiological dynamics in demyelinating diseases:unraveling complex relationships through computer modeling.Int J Mol Sci 16:21215-21236.
Du Z,Bondarenko O,Wang D,Rouabhia M,Zhang Z(2016)Ex vivo assay of electrical stimulation to rat sciatic nerves:cell behaviors and growth factor expression.J Cell Physiol 231:1301-1312.
Espinosa-Hoyos D,Jagielska A,Homan KA,Du H,Busbee T,Anderson DG,Fang NX,Lewis JA,Van Vliet KJ.(2018)Engineered 3D-printed artificial axons.Sci Rep 8:478.
Fields RD,Neale EA,Nelson PG.(1990)Effects of patterned electrical activity on neurite outgrowth from mouse sensory neurons.J Neurosci10:2950-2964.
Fields RD(2015)A new mechanism of nervous system plasticity:activity-dependent myelination.Nat Rev Neurosci 16:756-767
Fields RD(2014)Myelin formation and remodeling.Cell 156(1-2):15-17.
Geren BB,Schmitt FO(1954)The structure of the Schwann cell and its relation to the axon in certain invertebrate nerve fibers.Proc Natl Acad Sci U S A40:863-870.
Gertz CC,Leach MK,Birrell LK,Martin DC,Feldman EL,Corey JM(2010)Accelerated neuritogenesis and maturation of primary spinal motor neurons in response to nanofibers.Dev Neurobiol 70:589-603.
Howe CL(2006)Coated glass and vicryl microfibers as artificial axons.Cells Tissues Organs 183:180-194.
Ishibashi T,Dakin KA,Stevens B,Lee PR,Kozlov SV,Stewart CL,Fields RD(2006)Astrocytes promote myelination in response to electrical stimuluss.Neuron 49:823-832.
Lee HU,Blasiak A,Agrawal DR,Loong DTB,Thakor NV,All AH,Ho JS,Yang IH(2017)Subcellular electrical stimulation of neurons enhances the myelination of axons by oligodendrocytes.PLoS One 12:e0179642.
Lee S,Leach MK,Redmond SA,Chong SY,Mellon SH,Tuck SJ,Feng ZQ,Corey JM,Chan JR(2012)A culture system to study oligodendrocyte myelination processes using engineered nanofibers.Nat Methods 9:917-922.
Merolli A,Mao Y,Kohn J(2017)A suspended carbon fiber culture to model myelination by human Schwann cells.J Mater Sci Mater Med 28:57.
Nguyen KP,O’Neal TJ,Bolonduro OA,White E,Kravitz AV(2016)Feeding Experimentation Device(FED):A flexible open-source device for measuring feeding behavior.J Neurosci Methods 267:108-114.
Salzer JL(2015)Schwann cell myelination.Cold Spring Harb Perspect Biol7:a020529.
Schubert TW,D’Ausilio A,Canto R(2013)Using Arduino microcontroller boards to measure response latencies.Behav Res Methods 45:1332-1346.
Schultz BG,van Vugt FT(2016)Tap Arduino:An Arduino microcontroller for low-latency auditory feedback in sensorimotor synchronization experiments.Behav Res Methods 48:1591-1607.
Sheinin A,Lavi A,Michaelevski I(2015)StimDuino:an Arduino-based electrophysiological stimulus isolator.J Neurosci Methods 243:8-17.
Snaidero N,Simons M(2014)Myelination at a glance.J Cell Sci127(Pt14):2999-3004.
Stevens B,Tanner S,Fields RD(1998)Control of myelination by specific patterns of neural stimuluss.J Neurosci 18:9303-9311.
Subramanian A,Krishnan UM,Sethuraman S(2012)Axially aligned electrically conducting biodegradable nanofibers for neural regeneration.JMater Sci Mater Med 23:1797-1809.
Whitwam JG(1976)Classification of peripheral nerve fibres.An historical perspective.Anaesthesia 31:494-503.
Wilson MT,Goodwin DP,Brownjohn PW,Shemmell J,Reynolds JN.(2014)Numerical modelling of plasticity induced by transcranial magnetic stimulation.J Comput Neurosci 36:499-514.
Wu H,Ghekiere H,Beeckmans D,Tambuyzer T,van Kuyck K,Aerts JM,Nuttin B(2015)Conceptualization and validation of an open-source closed-loop deep brain stimulation system in rat.Sci Rep 4:9921.
Wu Y,Wang L,Guo B,Shao Y,Ma PX(2016)Electroactive biodegradable polyurethane significantly enhanced Schwann cells myelin gene expression and neurotrophin secretion for peripheral nerve tissue engineering.Biomaterials 87:18-31.
Wu Y,Wang L,Hu T,Ma PX,Guo B(2018)Conductive micropatterned polyurethane films as tissue engineering scaffolds for Schwann cells and PC12 cells.J Colloid Interface Sci 518:252-262.
Zalc B,Fields RD(2000)Do action potentials regulate myelination?Neuroscientist 6:5-13.
Zhang K,Zheng H,Liang S,Gao C(2016)Aligned PLLA nanofibrous scaffolds coated with graphene oxide for promoting neural cell growth.Acta Biomater 37:131-142.
Bechler ME,Byrne L,Ffrench-Constant C(2015)CNS Myelin Sheath Lengths Are an Intrinsic Property of Oligodendrocytes.Curr Biol25:2411-2416.
Bechler ME,Swire M,Ffrench-Constant C(2018)Intrinsic and adaptive myelination-A sequential mechanism for smart wiring in the brain.Dev Neurobiol 78:68-79.
Bullock PN,Rome LH(1990)Glass micro-fibers:a model system for study of early events in myelination.J Neurosci Res 27:383-393.
Buonanno A,Fields RD(1999)Gene regulation by patterned electrical activity during neural and skeletal muscle development.Curr Opin Neurobiol9:110-120.
Chang KJ,Redmond SA,Chan JR(2016)Remodeling myelination:implications for mechanisms of neural plasticity.Nat Neurosci 19:190-197.
Coggan JS,Bittner S,Stiefel KM,Meuth SG,Prescott SA(2015)Physiological dynamics in demyelinating diseases:unraveling complex relationships through computer modeling.Int J Mol Sci 16:21215-21236.
Du Z,Bondarenko O,Wang D,Rouabhia M,Zhang Z(2016)Ex vivo assay of electrical stimulation to rat sciatic nerves:cell behaviors and growth factor expression.J Cell Physiol 231:1301-1312.
Espinosa-Hoyos D,Jagielska A,Homan KA,Du H,Busbee T,Anderson DG,Fang NX,Lewis JA,Van Vliet KJ.(2018)Engineered 3D-printed artificial axons.Sci Rep 8:478.
Fields RD,Neale EA,Nelson PG.(1990)Effects of patterned electrical activity on neurite outgrowth from mouse sensory neurons.J Neurosci10:2950-2964.
Fields RD(2015)A new mechanism of nervous system plasticity:activity-dependent myelination.Nat Rev Neurosci 16:756-767
Fields RD(2014)Myelin formation and remodeling.Cell 156(1-2):15-17.
Geren BB,Schmitt FO(1954)The structure of the Schwann cell and its relation to the axon in certain invertebrate nerve fibers.Proc Natl Acad Sci U S A40:863-870.
Gertz CC,Leach MK,Birrell LK,Martin DC,Feldman EL,Corey JM(2010)Accelerated neuritogenesis and maturation of primary spinal motor neurons in response to nanofibers.Dev Neurobiol 70:589-603.
Howe CL(2006)Coated glass and vicryl microfibers as artificial axons.Cells Tissues Organs 183:180-194.
Ishibashi T,Dakin KA,Stevens B,Lee PR,Kozlov SV,Stewart CL,Fields RD(2006)Astrocytes promote myelination in response to electrical stimuluss.Neuron 49:823-832.
Lee HU,Blasiak A,Agrawal DR,Loong DTB,Thakor NV,All AH,Ho JS,Yang IH(2017)Subcellular electrical stimulation of neurons enhances the myelination of axons by oligodendrocytes.PLoS One 12:e0179642.
Lee S,Leach MK,Redmond SA,Chong SY,Mellon SH,Tuck SJ,Feng ZQ,Corey JM,Chan JR(2012)A culture system to study oligodendrocyte myelination processes using engineered nanofibers.Nat Methods 9:917-922.
Merolli A,Mao Y,Kohn J(2017)A suspended carbon fiber culture to model myelination by human Schwann cells.J Mater Sci Mater Med 28:57.
Nguyen KP,O’Neal TJ,Bolonduro OA,White E,Kravitz AV(2016)Feeding Experimentation Device(FED):A flexible open-source device for measuring feeding behavior.J Neurosci Methods 267:108-114.
Salzer JL(2015)Schwann cell myelination.Cold Spring Harb Perspect Biol7:a020529.
Schubert TW,D’Ausilio A,Canto R(2013)Using Arduino microcontroller boards to measure response latencies.Behav Res Methods 45:1332-1346.
Schultz BG,van Vugt FT(2016)Tap Arduino:An Arduino microcontroller for low-latency auditory feedback in sensorimotor synchronization experiments.Behav Res Methods 48:1591-1607.
Sheinin A,Lavi A,Michaelevski I(2015)StimDuino:an Arduino-based electrophysiological stimulus isolator.J Neurosci Methods 243:8-17.
Snaidero N,Simons M(2014)Myelination at a glance.J Cell Sci127(Pt14):2999-3004.
Stevens B,Tanner S,Fields RD(1998)Control of myelination by specific patterns of neural stimuluss.J Neurosci 18:9303-9311.
Subramanian A,Krishnan UM,Sethuraman S(2012)Axially aligned electrically conducting biodegradable nanofibers for neural regeneration.JMater Sci Mater Med 23:1797-1809.
Whitwam JG(1976)Classification of peripheral nerve fibres.An historical perspective.Anaesthesia 31:494-503.
Wilson MT,Goodwin DP,Brownjohn PW,Shemmell J,Reynolds JN.(2014)Numerical modelling of plasticity induced by transcranial magnetic stimulation.J Comput Neurosci 36:499-514.
Wu H,Ghekiere H,Beeckmans D,Tambuyzer T,van Kuyck K,Aerts JM,Nuttin B(2015)Conceptualization and validation of an open-source closed-loop deep brain stimulation system in rat.Sci Rep 4:9921.
Wu Y,Wang L,Guo B,Shao Y,Ma PX(2016)Electroactive biodegradable polyurethane significantly enhanced Schwann cells myelin gene expression and neurotrophin secretion for peripheral nerve tissue engineering.Biomaterials 87:18-31.
Wu Y,Wang L,Hu T,Ma PX,Guo B(2018)Conductive micropatterned polyurethane films as tissue engineering scaffolds for Schwann cells and PC12 cells.J Colloid Interface Sci 518:252-262.
Zalc B,Fields RD(2000)Do action potentials regulate myelination?Neuroscientist 6:5-13.
Zhang K,Zheng H,Liang S,Gao C(2016)Aligned PLLA nanofibrous scaffolds coated with graphene oxide for promoting neural cell growth.Acta Biomater 37:131-142.