Skin Bioprinting Protocol

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skin bioprinting protocol

Overview

Skin engineering is a prominent alternative to traditional methods used for wound healing, drug testing & discovery, and other skin-related applications. However, challenges remain when it comes to generating complex skin substitutes.  Follow this step-by-step protocol for skin bioprinting.

Collagen is extremely difficult to pattern in optimal concentrations (3 mg/mL).  With Allevi’s exclusive CORE™ printhead and this skin bioprinting protocol, you are now able to print and pattern pure 3 mg/mL type I collagen or 8 mg/mL methacrylated collagen. This is the first time that such low concentration collagen can be printed and patterned through 3D bioprinting. Allevi’s skin bioprinting protocol presents a novel solution to automating and standardizing the creation of skin models.

Materials

  1. Allevi printing dish
  2. 10 mL of Advanced Biomatrix PureCol
  3. Allevi 5 mL syringe
  4. 2 x Allevi layering tips™
  5. Syringe coupler
  6. NaOH
  7. 10X PBS
  8. Human neonatal dermal fibroblasts or human adult dermal fibroblasts
    • Note: you may also add keratinocytes and melanocytes
  9. Fibroblast growth medium

Methods

  1. Follow this protocol to prepare PureCol® or this protocol to prepare PhotoCol® and your cells for printing;
  2. Set your Allevi extruder to 4˚C;
  3. Attach the Allevi layering tip™ to your syringe;
  4. Bioprint on the Allevi printing dish;
    1. If you have an A3, set your bed plate to 37˚C to crosslink your skin 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
50.20.227
Pressure (PSI) Crosslink (sec)Print Temp (°C)
104

We hope that you found this step-by-step protocol for skin bioprinting useful! You are now ready to image and analyze your 3D bioprinted skin construct – click here for a full list of analysis protocols.

References

Lee, V., Singh, G., Trasatti, J. P., Bjornsson, C., Xu, X., Tran, T. N., … Karande, P. (2014). Design and Fabrication of Human Skin by Three-Dimensional Bioprinting. Tissue Engineering Part C: Methods, 20(6), 473–484. https://doi.org/10.1089/ten.tec.2013.0335

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