We highly recommend using the FRESH method for printing Lifeink® 200. FRESH printing gives you the ability to print high resolution, complex structures or tissues, without having to fill your ink with non-native materials such as alginate or cellulose.
Yes. Lifeink® 200 is a concentrated 35 mg/ml type I collagen bioink, comprised of ~97% type I collagen and ~3% type III collagen.
Yes - The directions for use document provides videos showing how to mix in extra media, acid, proteins, cells, or anything else to Lifeink® 200.
There are various methods to crosslink your bioprinted constructs.
For acellular constructs, there are more options. You can use chemical crosslinking via glutaraldehyde or genipin, or photocrosslinking with riboflavin.
For cellular constructs, you can add Ruthenium photoinitiator to the media that your construct is suspended in post print and photocrosslink.
You can also mix in other crosslinkable components to Lifeink prior to printing, such as alginate, methacrylated gelatin, PEGDA, or other tunable materials.
The method we recommend the most is letting the cells remodel, modify, and crosslink the native collagen matrix.
After the print is complete, simply incubate your dish containing the FRESH slurry/encapsulated printed construct at 37°C. The FRESH gelatin microparticles will melt, and your collagen will polymerize.
Once the collagen structure is released and floating in the melted gelatin, simply replace the gelatin with warm cell culture media. For example, pipette out 2 mL of melted slurry and replace with 2 mL of media. Repeat until the gelatin is fully washed out.