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
- 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
- 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.
- 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.
- Adjust pH of solution from step 2 to 7.4.
- Filter solution from step 3 with a 0.45 µm syringe filter under sterile conditions.*
- 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.)
- Mix in desired cell concentration in a sterile container. *
- Load cell-laden bioink into a 10 ml sterile luer lok syringe.
- Load syringe into BioBots printer.
- Prepare your design file through Repetier Host, then upload the sliced gcode file to biobots software.
* For prints without cells, step 4 may be skipped.
|Speed (mm/s)||Layer height (mm)||Nozzle Diam (mm)||Gauge|
|Pressure (psi)||Print Temp (°C)|
- Gelation time and gel stiffness can be adjusted by varying the concentration of PEGDA or LAP. For help adjusting print parameters please contact firstname.lastname@example.org.
- A fill volume change of more than 2 ml may affect pressure settings.
- 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
for further questions or information about these products, please contact email@example.com
- 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.
- Chan V et al. “Three-dimensional photopatterning of hydrogels using stereolithography for long-term cell encapstulation.” Lab Chip. 2010(10) 2062-70.
- 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.