Name: Title Cell Encapsulation by Droplets (Video) | JoVE Journal
Uploaded: 2007-10-01T13:09:54.000+00:00
Duration: 13 min 10 s
Description: Learn cell encapsulation by droplets for 3D bioprinting using a solenoid valve system. This method enables precise cell ejection and supports tissue engineering applications.

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NIH3T3 cells preparation:
A. Cells for Ejection
<ol>
<li>Trypsinize cells, then dilute 1:1 with cell media, and transfer from a T75 flask to a 15 mL Falcon tube</li>
<li>Spin down cells into a pellet by centrifuging, aspirate supernatant and wash cells with DPBS</li>
<li>Spin down cells into a pellet again, and aspirate supernatant</li>
<li>Resuspend cells in media</li>
<li>Determine cell density with hemocytometer (~200 X 104 cells/mL per T75 flask)</li>
<li>Centrifuge cell solution, aspirate supernatant, and resuspend in appropriate amount of media for varying cell concentrations</li>
</ol>
B. Cell ejection
<ol>
<li>Vortex cells before using for ejection</li>
<li>Transfer 200 &micro;L of cell solution into syringe</li>
<li>Set appropriate mode on pulse generator<ol>
<li>For ejecting single droplets and multiple droplets (bursts), set pulse generator to "E. BUR" mode</li>
<li>For continuous droplet ejection, set pulse generator to "NORM" mode</li>
</ol></li>
<li>Change signal settings<ol>
<li>Set high level and low level output voltage: HIL to 5 V and LOL to 0 V and make sure the"LIM"LED is on</li>
<li>Set signal as a square pulse</li>
<li>Change the amount of time the solenoid valve is open for droplet ejection by changing the value for "WID" or changing duty cycle ("DUTY")</li>
<li>Change the frequency of ejection by changing the value for "PER"</li>
<li>Change the number of droplets ejected in a burst by changing the value for "BUR"</li>
</ol></li>
<li>Eject cell solution onto prepared substrate for imaging with microscope</li>
</ol>
C. Staining
<ol>
<li>Make up dye solution with 0.5 &micro;L calcein-AM and 2 &micro;L ethidium homodimer per mL of DPBS</li>
<li>Immerse prepared substrate in dye solution</li>
<li>Allow sample to incubate for 10 minutes at 37&deg;C before imaging</li>
</ol>
Experiment Validation
<ol>
<li>On a Nikon Eclipse TE-2000 U Fluorescent Microscope<ol>
<li>Spot advanced software (Diagnostics, Inc.)</li>
<li>Live/Dead Assay</li>
</ol></li>
</ol>

The authors have nothing to disclose.

<table><tbody><tr><td>Fluorescent Microscope</td><td></td><td>Nikon Instruments</td><td>Eclipse TE-2000 U</td><td></td></tr><tr><td>Solenoid valve cell ejector</td><td></td><td></td><td></td><td>Operation frequency: 1 KHz, 30psi, 50nl~0.5ul. 12V Valve Driver: 2.5 Amp drive current</td></tr><tr><td>5-Gallon Portable air tank</td><td></td><td>Coleman</td><td>Powermate CT5</td><td>Pressurized air: 30 PSI </td></tr><tr><td>Pressure regulators</td><td></td><td>Marsh Bellofram</td><td></td><td></td></tr><tr><td>Pulse Generator</td><td></td><td></td><td>HP8112A</td><td>Actuation frequency generation: 50 MHz</td></tr><tr><td>XYZ-stage</td><td></td><td>Newmark Systems</td><td></td><td>With Stepping motor: 6inch travel xy-stage(NLS4-6-25), 2.5inch travel z-stage(NLS4-2.5-25), 3axis controller(NSC-G3), 0.1um resolution</td></tr><tr><td>NIH 3T3</td><td>Cell-line</td><td></td><td></td><td>fibroblasts</td></tr><tr><td>Trypsin</td><td></td><td></td><td></td><td>0.05% solution</td></tr><tr><td>NIH 3T3 cell medium</td><td></td><td></td><td></td><td></td></tr><tr><td>DPBS</td><td>Buffer</td><td></td><td></td><td></td></tr><tr><td>T75</td><td></td><td></td><td></td><td>Tissue culture flasks</td></tr><tr><td>Plastic conical tubes</td><td></td><td></td><td></td><td>15 ml, for tissue culture</td></tr></tbody></table>

title cell encapsulation by droplets

Title Cell Encapsulation by Droplets

My name is Hang Moon. I'm working with the professor of condoms. He and me work with the XYJ stage to make a 3D structure of cell.Hi, I am PE and Lynn and I'm working with the cell printing project with cell encapsulation and media and gels. This is solenoid be for ejection of a cell and cell mixture of aros over cell media. The so valve is activated by two signal line.The signal line is came from a dry board here. Signal came from here. That line through the that line, that one is the function first generator.So first is generated by our programmed way. So if you want, you want to change the purse, you can change the program with the that switches line or you can control that process generator to computer. The cellular valve is open and closed by the electric signal, but the, the cell is supplied through that line.The cell and the media or agro gel is supplied through the syringe syringes. The syringes inside of the water, inside of syringe is the press light by the air. The pressurized air is supplied from a tank like here the line.So the little line and the ized air is fly through the air filter. The air filter is discriminate the dust and other kind of the dirt. That is pressur

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Learn cell encapsulation by droplets for 3D bioprinting using a solenoid valve system. This method enables precise cell ejection and supports tissue engineering applications.

Title Cell Encapsulation by Droplets

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