Name: a multi-compartment cns neuron-glia co-culture microfluidic platform
Uploaded: 2009-09-10T16:30:00
Description: Watch this Scientific Journal Video about A Multi-compartment CNS Neuron-glia Co-culture Microfluidic Platform at JoVE.com

यह काम स्वास्थ्य / मानसिक स्वास्थ्य के राष्ट्रीय संस्थान (NIH / NIMH) के राष्ट्रीय संस्थानों 1R21MH085267 # अनुदान द्वारा समर्थित किया गया था.

भाग 1: बहु डिब्बे के लिए एक मास्टर मोल्ड तैयारी न्यूरॉन उपकरण निर्माण सह संस्कृति <ol><li> PMMA मास्टर बनाना पहले कदम के लिए 3.5 मिमी गहरी डिब्बों बनाना है. एक सोम डिब्बे और छह अक्षतंतु / glia डिब्बों एक PMMA एक सूक्ष्म ?...

<ol>
	<li>Sherman, D. L., Brophy, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+axon+ensheathment+and+myelin+growth.">Mechanisms of axon ensheathment and myelin growth.</a> Nature Reviews. 6, 683-690 (2005).</li><li>Xia, Y., Whitesides, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soft+Lithography.">Soft Lithography.</a> Angew. Chem. 37, 550-575 (1998).</li><li>Taylor, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+culture+platform+for+neuroscience+research.">Microfluidic culture platform for neuroscience research.</a> Nature Methods. 2, 599-605 (2005).</li><li>Li, J., Baud, O., Vartanian, T., Volpe, J. J., Rosenberg, P. A., A, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peroxynitrite+generated+by+inducible+nitric+oxide+synthase+and+NADPH+oxidase+mediates+microglial+toxicity+to+oligodendrocytes.">Peroxynitrite generated by inducible nitric oxide synthase and NADPH oxidase mediates microglial toxicity to oligodendrocytes.</a> Proceedings of the National Academy of Sciences. 102, 9936-9941 (2005).</li></ol>

हम स्तनधारी सीएनएस अक्षतंतु - glia बातचीत के अध्ययन के लिए एक बहु डिब्बे सह संस्कृति मंच विकसित किया है. सीएनएस axons सफलतापूर्वक neuronal सेल निकायों / dendrites से अलग थे, और oligodendrocytes सफलतापूर्वक थे डिवाइस के अंदर सह सुसं?...

a multi-compartment cns neuron-glia co-culture microfluidic platform

We present a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research, capable of conducting up to six independent experiments in parallel for higher-throughput. We developed a new fabrication method to create microfluidic devices having both micro and macro scale structures within the same device through a single soft-lithography process, enabling mass fabrication with good repeatability.
The multi-compartment microfluidic co-culture platform is composed of one soma compartment for neurons and six axon/glia compartments for oligodendrocytes (OLs). The soma compartment and axon/glia compartments are connected by arrays of axon-guiding microchannels that function as physical barriers to confine neuronal soma in the soma compartment, while allowing axons to grow into axon/glia compartments. OLs loaded into axon/glia compartments can interact only with axons but not with neuronal soma or dendrites, enabling localized axon-glia interaction studies. The microchannels also enabled fluidic isolation between compartments, allowing six independent experiments to be conducted on a single device for higher throughput.
Soft-lithography using poly(dimethylsiloxane) (PDMS) is a commonly used technique in biomedical microdevices. Reservoirs on these devices are commonly defined by manual punching. Although simple, poor alignment and time consuming nature of the process makes this process not suitable when large numbers of reservoirs have to be repeatedly created. The newly developed method did not require manual punching of reservoirs, overcoming such limitations. First, seven reservoirs (depth: 3.5 mm) were made on a poly(methyl methacrylate) (PMMA) block using a micro-milling machine. Then, arrays of ridge microstructures, fabricated on a glass substrate, were hot-embossed against the PMMA block to define microchannels that connect the soma and axon/glia compartments. This process resulted in macro-scale reservoirs (3.5 mm) and micro-scale channels (2.5 &mu;m) to coincide within a single PMMA master. A PDMS replica that served as a mold master was obtained using soft-lithography and the final PDMS device was replicated from this master.
Primary neurons from E16-18 rats were loaded to the soma compartment and cultured for two weeks. After one week of cell culture, axons crossed microchannels and formed axonal only network layer inside axon/glia compartments. Axons grew uniformly throughout six axon/glia compartments and OLs from P1-2 rats were added to axon/glia compartments at 14 days in vitro for co-culture.

Watch this Scientific Journal Video about A Multi-compartment CNS Neuron-glia Co-culture Microfluidic Platform at JoVE.com

A Multi-compartment CNS Neuron-glia Co-culture Microfluidic Platform

हम इन विट्रो सीएनएस बातचीत अक्षतंतु glia अनुसंधान में के लिए एक उपन्यास बहु - डिब्बे न्यूरॉन सह संस्कृति माइक्रोसिस्टम मंच विकसित की है. मंच समानांतर में छह स्वतंत्र प्रयोगों का आयोजन करने में सक्षम है और एक नव विकसित / मैक्रो सूक्ष्म संकर निर्माण विधि का उपयोग कर गढ़े.

एक डिब्बे मल्टी सीएनएस न्यूरॉन - सह - संस्कृति glia Microfluidic प्लेटफार्म

We present a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research, capable of conducting up to ...

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