Low density cell culture of locust neurons in closed-channel microfluidic devices.

Microfluidic channel systems were fabricated out of polydimethylsiloxane (PDMS) and used as culture vessels for primary culture of neurons from locust thoracic ganglia. In a biocompatibility study it was shown that cell adhesion and neuronal cell growth of locust neurons on uncoated PDMS was restricted. Coating with concanavalin A improved cell adhesion. In closed-channel microfluidic devices neurons were grown in static-bath culture conditions for more than 15 days. Cell densities of up to 20 cells/channel were not exceeded in low-density cultures but we also found optimal cell growth of single neurons inside individual channels. The first successful cultivation of insect neurons in closed-channel microfluidic devices provides a prerequisite for the development of low density neuronal networks on multi electrode arrays combined with microfluidic devices.

[1]  G. Whitesides,et al.  Poly(dimethylsiloxane) as a material for fabricating microfluidic devices. , 2002, Accounts of chemical research.

[2]  R. Campenot Local control of neurite sprouting in cultured sympathetic neurons by nerve growth factor. , 1987, Brain research.

[3]  P. Bräunig,et al.  Primary neuronal culture of Locusta migratoria for construction of networks on microelectronic recording devices , 2006 .

[4]  P. Usherwood,et al.  The effects of putative amino acid neurotransmitters on somata isolated from neurons of the locust central nervous system. , 1985, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[5]  Amir Ayali,et al.  A two-phase growth strategy in cultured neuronal networks as reflected by the distribution of neurite branching angles. , 2005, Journal of neurobiology.

[6]  Amir Ayali,et al.  The regulative role of neurite mechanical tension in network development. , 2009, Biophysical journal.

[7]  Amir Ayali,et al.  One-to-one neuron–electrode interfacing , 2009, Journal of Neuroscience Methods.

[8]  M. Pener,et al.  The physiology of locust phase polymorphism: an update. , 1998, Journal of insect physiology.

[9]  G. Banker,et al.  An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. II. Synaptic relationships , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[10]  David J. Beebe,et al.  Insect Cell Culture in Microfluidic Channels , 2002 .

[11]  Stefan Weigel,et al.  A simple fluorescent double staining method for distinguishing neuronal from non-neuronal cells in the insect central nervous system , 2006, Journal of Neuroscience Methods.

[12]  A. Offenhäusser,et al.  Ultrathin Coatings with Change in Reactivity over Time Enable Functional In Vitro Networks Of Insect Neurons , 2008, Advanced materials.

[13]  P. Bräunig,et al.  Neuronal cell growth on iridium oxide. , 2010, Biomaterials.

[14]  R. Lakes-Harlan,et al.  Thoracic Interneurons, Motorneurons and Sensory Neurons of Locusta Migratoria (Insecta: Orthoptera) in Primary Cell Culture , 2008 .

[15]  D. J. Beadle,et al.  Insect neuronal cultures: an experimental vehicle for studies of physiology, pharmacology and cell interactions , 2006, Invertebrate Neuroscience.

[16]  P. Usherwood,et al.  Locust nymphal neurones in culture: a new technique for studying the physiology and pharmacology of insect central neurones. , 1985, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[17]  G. Whitesides,et al.  Compatibility of mammalian cells on surfaces of poly(dimethylsiloxane). , 2004, Langmuir : the ACS journal of surfaces and colloids.

[18]  R. Nuzzo,et al.  Microfluidic devices for culturing primary mammalian neurons at low densities. , 2007, Lab on a chip.

[19]  Amir Ayali,et al.  Growth morphology of two-dimensional insect neural networks , 2002, Neurocomputing.

[20]  R. Campenot,et al.  Local control of neurite development by nerve growth factor. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[21]  E Claverol-Tinturé,et al.  Multielectrode arrays with elastomeric microstructured overlays for extracellular recordings from patterned neurons , 2005, Journal of neural engineering.

[22]  Long-term stability of PDMS-based microfluidic systems used for biocatalytic reactions , 2006 .

[23]  Bruce C Wheeler,et al.  Novel MEA platform with PDMS microtunnels enables the detection of action potential propagation from isolated axons in culture. , 2009, Lab on a chip.

[24]  M. Schachner,et al.  Spatiotemporal pattern of expression of tenascin-like molecules in a developing insect olfactory system. , 1994, Journal of neurobiology.

[25]  B Kirchhof,et al.  Growth properties of larval and adult locust neurons in primary cell culture , 1992, The Journal of comparative neurology.

[26]  Michael Kunst,et al.  In vivo labeling and in vitro characterisation of central complex neurons involved in the control of sound production , 2009, Journal of Neuroscience Methods.

[27]  S. Laughlin,et al.  Visual Targeting of Forelimbs in Ladder-Walking Locusts , 2010, Current Biology.

[28]  M. Burrows The Neurobiology of an Insect Brain , 1996 .

[29]  M. Pener Locust Phase Polymorphism and its Endocrine Relations , 1991 .

[30]  G. Banker,et al.  An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. I. Cells which develop without intercellular contacts , 1984, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[31]  Gregory J. Brewer,et al.  Survival and growth of hippocampal neurons in defined medium at low density: advantages of a sandwich culture technique or low oxygen , 1989, Brain Research.

[32]  Carl W. Cotman,et al.  β-Amyloid Induces Local Neurite Degeneration in Cultured Hippocampal Neurons: Evidence for Neuritic Apoptosis , 1998, Neurobiology of Disease.

[33]  E. Hulata,et al.  Coemergence of regularity and complexity during neural network development , 2007, Developmental neurobiology.

[34]  Boris Hofmann,et al.  Iridium Oxide Microelectrode Arrays for In Vitro Stimulation of Individual Rat Neurons from Dissociated Cultures , 2009, Front. Neuroeng..

[35]  M. Schachner,et al.  In vitro analyses of neurite outgrowth indicate a potential role for tenascin-like molecules in the development of insect olfactory glomeruli. , 1994, Journal of neurobiology.