Name: electric-field-induced neural precursor cell differentiation in microfluidic devices
Uploaded: 2021-04-14T14:00:00
Description: Watch this Scientific Journal Video about Electric-Field-Induced Neural Precursor Cell Differentiation in Microfluidic Devices at JoVE.com

The authors thank Professor Tang K. Tang, Institute of Biomedical Sciences, Academia Sinica, for his assistance in providing mouse neural stem and progenitor cells (mNPCs). The authors also thank Professor Tang K. Tang and Ms. Ying-Shan Lee, for their valuable discussion on the differentiation of mNPCs.

1. Design and fabrication of the MOE chip
<ol>
	<li>Draw patterns for individual polymethyl methacrylate (PMMA) layers and the double-sided tape using appropriate software (Figure 1A, Table of Materials). Cut both the PMMA sheets and the double-sided tape with a CO2 laser machine scriber (Figure 1B).
	<ol>
		<li>Switch on the CO2 laser scriber and connect it to a personal computer. Open the designed pattern file using the...

<ol>
	<li>Chang, H. F., Lee, Y. S., Tang, T. K., Cheng, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulsed+DC+electric+field-induced+differentiation+of+cortical+neural+precursor+cells.">Pulsed DC electric field-induced differentiation of cortical neural precursor cells.</a> PLoS One. 11 (6), e0158133 (2016).</li><li>Li, S., Li, H., Wang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orientation+of+spiral+ganglion+neurite+extension+in+electrical+fields+of+charge-balanced+biphasic+pulses+and+direct+current+in+vitro.">Orientation of spiral ganglion neurite extension in electrical fields of charge-balanced biphasic pulses and direct current in vitro.</a> Hearing research. 267 (1-2), 111-118 (2010).</li><li>Rajnicek, A. M., Robinson, K. R., McCaig, C. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrotaxis+of+lung+cancer+cells+in+a+multiple-electric-field+chip.">Electrotaxis of lung cancer cells in a multiple-electric-field chip.</a> Biosensors and Bioelectronics. 24 (12), 3510-3516 (2009).</li><li>Jing, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+of+electrical+stimulation+with+different+electric-field+intensities+in+the+regulation+of+the+differentiation+of+PC12+cells.">Study of electrical stimulation with different electric-field intensities in the regulation of the differentiation of PC12 cells.</a> American Chemical Society Chemical Neuroscience. 10 (1), 348-357 (2019).</li><li>Guo, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-powered+electrical+stimulation+for+enhancing+neural+differentiation+of+mesenchymal+stem+cells+on+graphene-poly(3,4-ethylenedioxythiophene)+hybrid+microfibers.">Self-powered electrical stimulation for enhancing neural differentiation of mesenchymal stem cells on graphene-poly(3,4-ethylenedioxythiophene) hybrid microfibers.</a> American Chemical Society Nano. 10 (5), 5086-5095 (2016).</li><li>Manabe, M., Mie, M., Yanagida, Y., Aizawa, M., Kobatake, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+effect+of+electrical+stimulation+and+cisplatin+in+HeLa+cell+death.">Combined effect of electrical stimulation and cisplatin in HeLa cell death.</a> Biotechnology and Bioengineering. 86 (6), 661-666 (2004).</li><li>Liu, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+effect+of+moderate+exogenous+electric+field+stimulation+on+activating+netrin-1/DCC+expression+against+mechanical+stretch-induced+injury+in+spinal+cord+neurons.">Protective effect of moderate exogenous electric field stimulation on activating netrin-1/DCC expression against mechanical stretch-induced injury in spinal cord neurons.</a> Neurotoxicity Research. 34 (2), 285-294 (2018).</li><li>Haan, N., Song, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+application+of+electric+fields+in+the+injured+nervous+system.">Therapeutic application of electric fields in the injured nervous system.</a> Advances in Wound Care. 3 (2), 156-165 (2014).</li><li>Ariza, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+electric+fields+on+hippocampal+neural+progenitor+cells.">The influence of electric fields on hippocampal neural progenitor cells.</a> Stem Cell Reviews and Reports. 6 (4), 585-600 (2010).</li><li>Huang, C. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+expression+of+human+lung+cancer+cell+line+CL1-5+in+response+to+a+direct+current+electric+field.">Gene expression of human lung cancer cell line CL1-5 in response to a direct current electric field.</a> PLoS One. 6 (10), e25928 (2011).</li><li>Sun, Y. S., Peng, S. W., Lin, K. H., Cheng, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrotaxis+of+lung+cancer+cells+in+ordered+three-dimensional+scaffolds.">Electrotaxis of lung cancer cells in ordered three-dimensional scaffolds.</a> Biomicrofluidics. 6 (1), 14102-14114 (2012).</li><li>Hou, H. S., Chang, H. F., Cheng, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrotaxis+studies+of+lung+cancer+cells+using+a+multichannel+dual-electric-field+microfluidic+chip.">Electrotaxis studies of lung cancer cells using a multichannel dual-electric-field microfluidic chip.</a> Journal of Visualized Experiments. 106, e53340 (2015).</li><li>Cheng, J. Y., Yen, M. H., Hsu, W. C., Jhang, J. H., Young, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ITO+patterning+by+a+low+power+Q-switched+green+laser+and+its+use+in+the+fabrication+of+a+transparent+flow+meter.">ITO patterning by a low power Q-switched green laser and its use in the fabrication of a transparent flow meter.</a> Journal of Micromechanics and Microengineering. 17 (11), 2316-2323 (2007).</li><li>Cheng, J. Y., Yen, M. H., Kuo, C. T., Young, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+transparent+cell-culture+microchamber+with+a+variably+controlled+concentration+gradient+generator+and+flow+field+rectifier.">A transparent cell-culture microchamber with a variably controlled concentration gradient generator and flow field rectifier.</a> Biomicrofluidics. 2 (2), 24105 (2008).</li><li>Fuhr, G., Shirley, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+handling+and+characterization+using+micron+and+submicron+electrode+arrays:+state+of+the+art+and+perspectives+of+semiconductor+microtools.">Cell handling and characterization using micron and submicron electrode arrays: state of the art and perspectives of semiconductor microtools.</a> Journal of Micromechanics and Microengineering. 5 (2), 77-85 (1995).</li><li>AChang, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biphasic+electrical+currents+stimulation+promotes+both+proliferation+and+differentiation+of+fetal+neural+stem+cells.">Biphasic electrical currents stimulation promotes both proliferation and differentiation of fetal neural stem cells.</a> PLoS One. 6 (4), e18738 (2011).</li><li>Lim, J. H., McCullen, S. D., Piedrahita, J. A., Loboa, E. G., Olby, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alternating+current+electric+fields+of+varying+frequencies:+effects+on+proliferation+and+differentiation+of+porcine+neural+progenitor+cells.">Alternating current electric fields of varying frequencies: effects on proliferation and differentiation of porcine neural progenitor cells.</a> Cell Reprogram. 15 (5), 405-412 (2013).</li></ol>

During the fabrication of the MOE chip, the adaptors are attached to the Layer 1 of the MOE chip with fast-acting cyanoacrylate glue. The glue is applied to 4 corners of the adaptors, and then pressure is applied evenly over the adaptors. Excess amount of glue must be avoided to ensure complete polymerization of the glue. Moreover, the completed MOE chip assembly is incubated in a vacuum chamber. This step helps to remove the bubbles between the PMMA layer, the double-sided tape, and the cover glass.
<p class="jove_c...

Neural precursor cells (NPCs, also known as neural stem and progenitor cells) can be as a promising candidate for neurodegenerative therapeutic strategy1. The undifferentiated NPCs have self-renewal capacity, multi-potency, and proliferative ability2,3. A previous study has reported that the extracellular matrix and molecular mediators regulate differentiation of NPC. The epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) promote NPC proliferation, thus maintaining the undifferentiated state4.
Previous studies have reported that electrical stimulation can regulate cell physiologic activities such as division5, migration6,7,8, differentiation1,9,10, and cell death11. Electric fields (EFs) play vital roles in the development and regeneration of the central nervous system development12,13,14. From 2009 to 2019, this laboratory has investigated cellular responses to the application of EF in the microfluidic system1,6,7,8,15,16,17. A multichannel, optically transparent, electrotactic (MOE) chip was designed to be suitable for immunofluorescence staining for confocal microscopy. The chip had high optical transparency and good durability and allowed the simultaneous conduct of three independent stimulation experiments and several immunostained conditions in a single study. The microfluidic system is beneficial in simplifying the overall experimental setup and, in turn, the reagent and sample consumption. This paper describes the development of a microfluidic cell culture system that was used for a long-term cell differentiation study.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mm PMMA substrates (Layers 1-3)</td>
			<td>BHT</td>
			<td>K2R20</td>
			<td>Polymethyl methacrylate (PMMA), http://www.bothharvest.com/zh-tw/product-421076/Optical-PMMA-Non-Coated-BHT-K2Rxx-xx=-thickness-choices.html</td>
		</tr>
		<tr>
			<td>15 mL plastic tube</td>
			<td>Protech Technology Enterprise Co., Ltd</td>
			<td>CT-15-PL-TW</td>
			<td>Conical bottomed tube with cap, assembled, presterilized</td>
		</tr>
		<tr>
			<td>3 mL syringe</td>
			<td>TERUMO</td>
			<td>DVR-3413</td>
			<td>3 mL oral syringes, without needle</td>
		</tr>
		<tr>
			<td>3 mm optical grade PMMA (Layer 5)</td>
			<td>CHI MEI Corporation</td>
			<td>ACRYPOLY PMMA Sheet</td>
			<td>Optical grade PMMA</td>
		</tr>
		<tr>
			<td>3-way stopcock</td>
			<td>NIPRO</td>
			<td>NCN-3L</td>
			<td>Sterile disposable 3-way stopcock</td>
		</tr>
		<tr>
			<td>5 mL syringe</td>
			<td>TERUMO</td>
			<td>DVR-3410</td>
			<td>5 mL oral syringes, without needle</td>
		</tr>
		<tr>
			<td>Adaptor</td>
			<td>Dong Zhong Co., Ltd.</td>
			<td>Customized</td>
			<td>PMMA adaptor</td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Sigma-Aldrich</td>
			<td>A9414</td>
			<td>Agarose, low gelling temperature</td>
		</tr>
		<tr>
			<td>Amplifier</td>
			<td>A.A. Lab Systems Ltd</td>
			<td>A-304</td>
			<td>High voltage amplifier</td>
		</tr>
		<tr>
			<td>AutoCAD software</td>
			<td>Autodesk</td>
			<td>Educational Version</td>
			<td>Drafting</td>
		</tr>
		<tr>
			<td>B-27 supplement</td>
			<td>Gibco</td>
			<td>12587-010</td>
			<td>B-27 supplement (50x), minus vitamin A</td>
		</tr>
		<tr>
			<td>Basic fibroblast growth factor (bFGF)&#160;</td>
			<td>Peprotech</td>
			<td>AF-100-18B</td>
			<td>Also called recombinant human FGF-basic</td>
		</tr>
		<tr>
			<td>Black rubber bung</td>
			<td>TERUMO</td>
			<td>DVR-3413</td>
			<td>From 3 mL oral syringes, without needle</td>
		</tr>
		<tr>
			<td>Bovine serum albumin (BSA)</td>
			<td>Sigma-Aldrich</td>
			<td>B4287</td>
			<td>Blocking reagent&#160;</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>HSIANGTAI</td>
			<td>CV2060</td>
			<td>Centrifuge</td>
		</tr>
		<tr>
			<td>CO2 laser scriber</td>
			<td>Laser Tools and Technics Corp.&#160;</td>
			<td>ILS-II</td>
			<td>Purchased from http://www.lttcorp.com/index.htm</td>
		</tr>
		<tr>
			<td>Cone connector</td>
			<td>IDEX Health &#38; Science</td>
			<td>F-120X</td>
			<td>One-piece fingertight 10-32 coned, for 1/16&#34; OD natural</td>
		</tr>
		<tr>
			<td>Cone-Luer adaptor</td>
			<td>IDEX Health &#38; Science</td>
			<td>P-659</td>
			<td>Luer Adapter 10-32 Female to Female Luer, PEEK</td>
		</tr>
		<tr>
			<td>Confocal fluorescence microscope</td>
			<td>Leica Microsystems</td>
			<td>TCS SP5</td>
			<td>Leica TCS SP5 user manual, http://www3.unifr.ch/bioimage/wp-content/uploads/2013/10/User-Manual_TCS_SP5_V02_EN.pdf</td>
		</tr>
		<tr>
			<td>Digital camera</td>
			<td>OLYMPUS</td>
			<td>E-330</td>
			<td>Automatic time-lapse image acquisition</td>
		</tr>
		<tr>
			<td>Digital oscilloscope</td>
			<td>Tektronix</td>
			<td>TDS2024</td>
			<td>Measure voltage or current signals over time in an electronic circuit or component to display amplitude and frequency.</td>
		</tr>
		<tr>
			<td>Double-sided tape</td>
			<td>3M&#160;</td>
			<td>PET 8018</td>
			<td>Purchased from http://en.thd.com.tw/</td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s modified Eagle&#8217;s medium/Ham&#39;s nutrient mixture F-12 (DMEM/F12)</td>
			<td>Gibco</td>
			<td>12400024</td>
			<td>DMEM/F-12, powder, HEPES</td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s phosphate-buffered saline (DPBS)</td>
			<td>Gibco</td>
			<td>21600010</td>
			<td>DPBS, powder, no calcium, no magnesium</td>
		</tr>
		<tr>
			<td>EF multiplexer</td>
			<td>Asiatic Sky Co., Ltd.</td>
			<td>Customized</td>
			<td>Monitor and control the electric current in individual channels</td>
		</tr>
		<tr>
			<td>Epidermal growth factor (EGF)</td>
			<td>Peprotech</td>
			<td>AF-100-15</td>
			<td>Also called recombinant human EGF</td>
		</tr>
		<tr>
			<td>Fast-acting cyanoacrylate glue</td>
			<td>3M&#160;</td>
			<td>7004T</td>
			<td>Strength instant adhesive (liquid)</td>
		</tr>
		<tr>
			<td>Flat bottom connector</td>
			<td>IDEX Health &#38; Science</td>
			<td>P-206</td>
			<td>Flangeless male nut Delrin, 1/4-28 flat-bottom, for 1/16&#34; OD blue</td>
		</tr>
		<tr>
			<td>Function generator</td>
			<td>Agilent Technologies</td>
			<td>33120A</td>
			<td>High-performance 15 MHz synthesized&#160;function generator&#160;with built-in arbitrary waveform capability</td>
		</tr>
		<tr>
			<td>Goat anti-mouse IgG H&#38;L (Alexa Fluor 488)</td>
			<td>Abcam</td>
			<td>ab150117</td>
			<td>Goat anti-mouse IgG H&#38;L (Alexa Fluor 488) preadsorbed</td>
		</tr>
		<tr>
			<td>Goat anti-rabbit IgG H&#38;L (Alexa Fluor 555)</td>
			<td>Abcam</td>
			<td>ab150086</td>
			<td>Goat polyclonal secondary antibody to rabbit IgG - H&#38;L (Alexa Fluor 555), preadsorbed</td>
		</tr>
		<tr>
			<td>Hoechst 33342</td>
			<td>Invitrogen</td>
			<td>H3570</td>
			<td>Nuclear staining</td>
		</tr>
		<tr>
			<td>ImageJ software</td>
			<td>National Institutes of Health</td>
			<td>1.48v</td>
			<td>Analyze the fluorescent images&#160;</td>
		</tr>
		<tr>
			<td>Indium&#8211;tin&#8211;oxide (ITO) glass</td>
			<td>Merck</td>
			<td>300739</td>
			<td>For ITO heater</td>
		</tr>
		<tr>
			<td>Inverted phase contrast microscope</td>
			<td>OLYMPUS</td>
			<td>CKX41</td>
			<td>For cell morphology observation</td>
		</tr>
		<tr>
			<td>K-type thermocouple</td>
			<td>Tecpel</td>
			<td>TPK-02A</td>
			<td>Temperature&#160;thermocouples</td>
		</tr>
		<tr>
			<td>Luer adapter</td>
			<td>IDEX Health &#38; Science</td>
			<td>P618-01</td>
			<td>Luer adapter female Luer to 1/4-28 male polypropylene</td>
		</tr>
		<tr>
			<td>Luer lock syringe</td>
			<td>TERUMO</td>
			<td>DVR-3413</td>
			<td>For agar salt bridges</td>
		</tr>
		<tr>
			<td>Mouse anti-GFAP</td>
			<td>eBioscience</td>
			<td>14-9892</td>
			<td>Astrocytes marker</td>
		</tr>
		<tr>
			<td>Oligodendrocyte &#160;marker&#160; O4&#160; antibody</td>
			<td>R&#38;D Systems</td>
			<td>MAB1326</td>
			<td>Oligodendrocytes marker</td>
		</tr>
		<tr>
			<td>Paraformaldehyde (PFA)</td>
			<td>Sigma-Aldrich</td>
			<td>P6148</td>
			<td>Fixing&#160;agent</td>
		</tr>
		<tr>
			<td>Phosphate buffered saline (PBS)</td>
			<td>Basic Life</td>
			<td>BL2651</td>
			<td>Washing solution</td>
		</tr>
		<tr>
			<td>Poly-L-Lysine (PLL)</td>
			<td>SIGMA</td>
			<td>P4707</td>
			<td>Coating solution</td>
		</tr>
		<tr>
			<td>Precision cover glasses thickness No. 1.5H</td>
			<td>MARIENFELD</td>
			<td>107242</td>
			<td>https://www.marienfeld-superior.com/precision-cover-glasses-thickness-no-1-5h-tol-5-m.html</td>
		</tr>
		<tr>
			<td>Programmable X-Y-Z motorised stage</td>
			<td>Tanlian Inc</td>
			<td>Customized</td>
			<td>Purchased from http://www.tanlian.tw/ndex.files/motort.htm</td>
		</tr>
		<tr>
			<td>Proportional&#8211;integral&#8211;derivative (PID) controller</td>
			<td>Toho Electronics</td>
			<td>TTM-J4-R-AB</td>
			<td>Temperature controller&#160;</td>
		</tr>
		<tr>
			<td>PTFE tube</td>
			<td>Professional Plastics Inc. Taiwan Branch</td>
			<td>Outer diameter 1/16 Inches</td>
			<td>White translucent PTFE tubing</td>
		</tr>
		<tr>
			<td>Rabbit anti-Tuj1</td>
			<td>Abcam</td>
			<td>ab18207</td>
			<td>Neuron marker</td>
		</tr>
		<tr>
			<td>Syringe pump</td>
			<td>New Era Systems Inc</td>
			<td>NE-1000</td>
			<td>NE-1000 programmable single syringe pump</td>
		</tr>
		<tr>
			<td>TFD4 detergent</td>
			<td>FRANKLAB</td>
			<td>TFD4</td>
			<td>Cover glass cleaner</td>
		</tr>
		<tr>
			<td>Thermal bonder</td>
			<td>Kuan-MIN Tech Co.</td>
			<td>Customized</td>
			<td>Purchased from http://kmtco.com.tw/</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma-Aldrich</td>
			<td>T8787</td>
			<td>Permeabilized solution</td>
		</tr>
		<tr>
			<td>Ultrasonic cleaner</td>
			<td>LEO</td>
			<td>LEO-300S</td>
			<td>Ultrasonic steri-cleaner</td>
		</tr>
		<tr>
			<td>Vacuum chamber</td>
			<td>DENG YNG INSTRUMENTS CO., Ltd.</td>
			<td>DOV-30</td>
			<td>Vacuum drying oven</td>
		</tr>
		<tr>
			<td>White fingertight plug</td>
			<td>IDEX Health &#38; Science</td>
			<td>P-316</td>
			<td>1/4-28 Flat-Bottom, https://www.idex-hs.com/store/fluidics/fluidic-connections/plug-teflonr-pfa-1-4-28-flat-bottom.html</td>
		</tr>
	</tbody>
</table>

electric-field-induced neural precursor cell differentiation in microfluidic devices

Physiological electric fields (EF) play vital roles in cell migration, differentiation, division, and death. This paper describes a microfluidic cell culture system that was used for a long-term cell differentiation study using microscopy. The microfluidic system consists of the following major components: an optically transparent electrotactic chip, a transparent indium-tin-oxide (ITO) heater, a culture media-filling pump, an electrical power supply, a high-frequency power amplifier, an EF multiplexer, a programmable X-Y-Z motorized stage, and an inverted phase-contrast microscope equipped with a digital camera. The microfluidic system is beneficial in simplifying the overall experimental setup and, in turn, the reagent and sample consumption. This work involves the differentiation of neural stem and progenitor cells (NPCs) induced by direct current (DC) pulse stimulation. In the stem cell maintenance medium, the mouse NPCs (mNPCs) differentiated into neurons, astrocytes, and oligodendrocytes after the DC pulse stimulation. The results suggest that simple DC pulse treatment could control the fate of mNPCs and could be used to develop therapeutic strategies for nervous system disorders. The system can be used for cell culture in multiple channels, for long-term EF stimulation, for cell morphological observation, and for automatic time-lapse image acquisition. This microfluidic system not only shortens the required experimental time, but also increases the accuracy of control on the microenvironment.

The detailed configuration of the MOE chip is shown in Figure 1. The microfluidic chip provides a beneficial approach for reducing the experimental setup size, sample volume, and reagent volume. The MOE chip was designed to perform three independent EF stimulation experiments and several immunostaining conditions simultaneously in a single study (Figure 3). In addition, the MOE chip, which has a high optical transparency is suita...

Watch this Scientific Journal Video about Electric-Field-Induced Neural Precursor Cell Differentiation in Microfluidic Devices at JoVE.com

Electric-Field-Induced Neural Precursor Cell Differentiation in Microfluidic Devices

In this study, we present a protocol for the differentiation of neural stem and progenitor cells (NPCs) solely induced by direct current (DC) pulse stimulation in a microfluidic system.

Physiological electric fields (EF) play vital roles in cell migration, differentiation, division, and death. This paper describes a microfluidic cell ...

Research

JoVE Journal

Bioengineering

Separating Beads and Cells in Multi-channel Microfluidic Devices Using Dielectrophoresis and Laminar Flow

Microfluidic devices have advanced cell studies by providing a dynamic fluidic environment on the scale of the cell for studying, manipulating, sorting ...

Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish

Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish

Micro fabricated fluidic devices provide an accessible micro-environment for in vivo studies on small organisms. Simple fabrication processes are ...

Detection of Exosomal Biomarker by Electric Field-induced Release and Measurement (EFIRM)

Detection of Exosomal Biomarker by Electric Field-induced Release and Measurement EFIRM

Exosomes are microvesicular structures that play a mediating role in intercellular communication. It is of interest to study the internal cargo of ...

Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization

Although microfluidics provides exquisite control of the cellular microenvironment, culturing cells within microfluidic devices can be challenging. 3D ...

Using Adhesive Patterning to Construct 3D Paper Microfluidic Devices

We demonstrate the use of patterned aerosol adhesives to construct both planar and nonplanar 3D paper microfluidic devices. By spraying an aerosol ...

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; ...

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to ...

Fabrication of Three-dimensional Paper-based Microfluidic Devices for Immunoassays

Paper wicks fluids autonomously due to capillary action. By patterning paper with hydrophobic barriers, the transport of fluids can be controlled and ...

Polydimethylsiloxane-polycarbonate Microfluidic Devices for Cell Migration Studies Under Perpendicular Chemical and Oxygen Gradients

This paper reports a microfluidic device made of polydimethylsiloxane (PDMS) with an embedded polycarbonate (PC) thin film to study cell migration under ...

Development of Microfluidic Devices to Study the Elongation Capability of Tip-growing Plant Cells in Extremely Small Spaces

In vivo, tip-growing plant cells need to overcome a series of physical barriers; however, researchers lack the methodology to visualize cellular behavior ...