Bioprinting Collagen for Layered Tissues

Overview

Collagen type I is the most abundant protein in the extracellular matrix and has been widely used in academia and industry for tissue engineering applications. With Allevi’s exclusive CORE™ printhead and this bioprinting collagen protocol, you are now able to print and pattern pure 3 mg/mL type I collagen or 8 mg/mL methacrylated collagen for layered tissues such as skin. This is the first time that such low concentration collagen can be printed and patterned through 3D bioprinting. 

Materials

• Advanced Biomatrix PureCol®
• Allevi 5 mL Syringe
• Allevi Layering Tips
• Syringe Coupler
• NaOH
• 10X PBS
• CytoSoft®
• Cell Media
• Cells

Methods 

1. Follow this protocol to prepare PureCol® and your cells for printing;
2. Set your Allevi extruder temperature to 4˚C;
3. Attach the Allevi layering tip™ to your syringe;
4. Bioprint on the Allevi printing dish;
   • If you have an A3, set your bed plate to 37˚C to crosslink your collagen layers at the time of deposition;
5. Incubate your structure at 37˚C for at least 30 minutes; 
6. Culture your layered tissue as desired.

Print Settings

Speed (mm/s)	Layer height (mm)	Nozzle Diam (mm)	Gauge
5	0.2	0.2	27

Pressure (PSI)	Crosslink (sec)	Print Temp (°C)
10	–	4

We hope you found this protocol for bioprinting collagen for layered tissues helpful! You are now ready for imaging and analysis of your layered tissue construct – click here for analysis protocols.