Directions for Use

The DFU below is for Lifeink 240, though the exact same techniques and directions apply for the higher concentration version, Lifeink 260

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Note: Employ aseptic practices to maintain the sterility of the product throughout the preparation and handling of the collagen and other solutions.

Note: Ensure that NO bubbles enter the system if mixing other materials. Bubbles in the system while mixing will turn your ink into a foam-like material.

Note: Cells should not be added directly to the Lifeink® 240 bioink since it is an acidic formulation.

Addition of other components to Lifeink® 260 Bioink:

Note: Components other than cells can be added to the Lifeink® 260 bioink before bioprinting as long as these components are compatible with acidic pH conditions.

1. To mix in additives to the bioink, add the additives to a secondary syringe as demonstrated in the video below:

Video for step 1

Note: The video shows cells being added to the bioink.  Other materials can be added to the Lifeink® 260 bioink in lieu of cells. Cells should NOT be added directly to this bioink. For cellular bioprinting, use Lifeink® 200 #5278-5ML.

Note: For best results, add no more than 5 mL of other components per 5 mL of collagen bioink. Use a similar ratio for smaller volumes.

2. Place sterile coupler on the end of the syringe with the bioink additives.
3. Slowly push plunger in until solution forms a slight external meniscus above the end of the coupler on the syringe.
4. Remove cap from the syringe with collagen and slowly push plunger in until collagen forms a slight external meniscus above the end of the syringe.
5. Couple the syringe with additives to the syringe with collagen. (Ensure that there are no air bubbles in the system. The “external meniscus” on both syringes helps ensure that there are no air bubbles introduced).
6. Slowly push plungers back and forth ~40 times to ensure thorough mixing. End with all of the material in the syringe to be used for printing.

Video for steps 2-6

7. The bioink with other components is now ready for extrusion 3D bioprinters.

Note: For pneumatic printers, transfer the collagen into an appropriate syringe using the coupler. The new syringe should have the seal inserted, but the plunger removed. Centrifuge the syringe at 2000 RPM for 1 minute after transferring the collagen to remove any air bubbles.

Watch this video for help

General Printing Notes:

1. To use a smaller volume of collagen, simply transfer the desired amount of collagen to another syringe, using the provided sterile coupler. To remove the air from the new syringe, you can do either of the following:

1. 1. Centrifuge the syringe (capped) with the cap pointing up to cause the air to accumulate at the cap. Evacuate the air.
   2. Centrifuge the syringe (capped) with the cap pointing down, and then use a hemostat to squeeze the syringe while pushing the plunger to allow the air to escape.

Removing air with a hemostat video

2. When printing with FRESH gelatin slurry, allow the final printed structure to incubate at 37°C for 30 to 60 minutes and then replace the gelatin with media.
3. Avoid bubbles.

For more directions on FRESH printing, please visit our LifeSupport® Directions for Use.

Product Applications

Read our Bioprinting eBrochure Here

Product Bioprints

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Product References

References for Lifeink^® collagen bioinks:

Lan, X. et al. In vitro maturation and in vivo stability of bioprinted human nasal cartilage. Journal of Tissue Engineering 13, 204173142210863 (2022).

Stocco, T. D., Moreira Silva, M. C., Corat, M. A., Gonçalves Lima, G. & Lobo, A. O. Towards bioinspired meniscus-regenerative scaffolds: Engineering A novel 3D bioprinted patient-specific construct reinforced by biomimetically aligned nanofibers. International Journal of Nanomedicine Volume 17, 1111–1124 (2022).

Lee, A. et al. 3D bioprinting of collagen to rebuild components of the human heart. Science 365, 482–487 (2019).

Maxson, Eva L., et al. "In vivo remodeling of a 3D-Bioprinted tissue engineered heart valve scaffold." Bioprinting (2019): e00059.

Filardo, G. et al.Patient-specific meniscus prototype based on 3D bioprinting of human cell-laden scaffold. Bone & Joint Research 8,101–106 (2019).

Schmitt, T. Analysis and Classification of 3-D Printed Collagen-Bioglass Matrices for Cellular Growth Utilizing Artificial Neural Networks. University Thesis (2018).

Balakhovsky, Y. M., Ostrovskiy, A. Y. & Khesuani, Y. D. Emerging Business Models Toward Commercialization of Bioprinting Technology. 3D Printing and Biofabrication1–22 (2017). doi:10.1007/978-3-319-40498-1_25-1

Fox, S. et al. A simplified fabrication technique for cellularized high-collagen dermal equivalents. Biomedical Materials14,041001 (2019).

Lin, H.-H., Chao, P.-H. G., Tai, W.-C. & Chang, P.-C. 3D-printed collagen-based waveform microfibrous scaffold for periodontal ligament reconstruction. International Journal of Molecular Sciences 22, 7725 (2021).

Engberg, A., Stelzl, C., Eriksson, O., O’Callaghan, P. & Kreuger, J. An open source extrusion bioprinter based on the E3D motion system and Tool Changer to enable fresh and multimaterial bioprinting. Scientific Reports 11, (2021).

Tashman, J., et al. "In Situ Volumetric Imaging and Analysis of FRESH 3D Bioprinted Constructs Using Optical Coherence Tomography." BioRxiv. (2021) 

Product Videos

Lifeink Collagen Bioink

Bioprinted Constructs Video

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Introduction to Lifeink

Lifeink Instructional Video

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Safety and Documentation

Safety Data Sheet

Certificate of Origin

Declaration of Material Source

Product Disclaimer

This product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.