PEGDA

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Overview

Bioprint_01_10X1X_200microns3D

Figure 1:  Live/dead image of human neonatal dermal fibroblasts encapsulated in bioprinted PEGDA

Poly(ethyelene glycol) diacryate is a PEG-based hydrogel that reacts and crosslinks in the presence of lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) and BioBots blue light technology(1,2). The recommended preparation provided in the user instruction below yields a streamlined matrix bioink that supports 3D printed cell-laden constructs. However, preparation can be modified by users to suit their needs.

note: due to low viscosity, PEGDA solutions can be difficult to work with when bioprinting. For optimal results, we suggest mixing this reagent with a more viscous material. 

Storage and Handling

PEGDA should be stored at -20 °C. LAP should be stored away from light in a dry environment.

You Will Need

  • PEGDA from BioBots
  • LAP from BioBots
  • Percoll
  • Metal or UV-block needles (we suggest these)
  •  Petri dish or well plate for printing
  •  0.45 µm Sterile filters (we suggest these)
  •  Optional: Treated glass (See protocol)

Instructions for Use

  1.  Measure 0.8% w/v LAP concentration and mix in cell media or PBS at 60°C in a light-protected vial. Mix until all LAP is dissolved.
  2.  Measure 33.33% w/v PEGDA concentration and add to solution from step 1, still in a light-protected vial. Mix at 60°C until all PEGDA is dissolved.
  3. Adjust pH of solution from step 2 to 7.4.
  4. Filter solution from step 3 with a 0.45 µm syringe filter under sterile conditions.*
  5. Mix solution from step 4 with Percoll to create a 40% (v/v) Percoll solution. (For 5 ml, you will need 3 ml of solution from step 4 and 2 ml of Percoll solution.)
  6. Mix in desired cell concentration in a sterile container. *
  7.  Load cell-laden bioink into a 10 ml sterile luer lok syringe.
  8.  Load syringe into BioBots printer.
  9.  Prepare your design file through Repetier Host, then upload the sliced gcode file to biobots software.
    1. Note that this material will need to be combined with some type of support or sacrificial reagent such as pluronic or PCL.

* For prints without cells, step 4 may be skipped.

Print Settings

Slic3r Settings
Speed (mm/s) Layer height (mm) Nozzle Diam (mm)   Gauge

4.0

0.3 0.3 30 tapered

Print Pressure and Temperature Settings
Pressure (psi) Print Temp (°C)

3-5

25
Print Crosslink Settings

Layers

 Intensity Time (sec)

0

0 0
Post Print Crosslink Settings

Intensity

Time (sec)

80%

240

Notes

  1.  Gelation time and gel stiffness can be adjusted by varying the concentration of PEGDA or LAP. For help adjusting print parameters please contact info@biobots.io.
  2.  A fill volume change of more than 2 ml may affect pressure settings.
  3.  A lower gauge size or tapered gauge will require lower pressure, while a higher gauge will require higher pressure for extrusion. Lowering the gauge size will also generally lower resolution

Further Information

BioKey

for further questions or information about these products, please contact info@biobots.io

References

  1. B. D. Fairbanks et. al, “Photoinitiated Polymerization of PEG-diacrylate with lithium phenyl-2,4,6-trimethylbenzoylphosphinate: polymerization rate and cytocompatibility,Biomaterials, vol. 30, no. 35, pp. 6702-6707, Dec 2009.
  2. Chan V et al. “Three-dimensional photopatterning of hydrogels using stereolithography for long-term cell encapstulation.” Lab Chip. 2010(10) 2062-70.
  3. Lin, Hang et al. “Application of Visible Light-based Projection Stereolithography for Live Cell-Scaffold Fabrication with Designed Architecture.” Biomaterials 2013(34). pp. 331- 39.