Instructions for Use

 Kraken Collagen Hydrogel 18 mg/mL- Research Use Only

Intended Use

This Instructions for Use document contains information on how to prepare and extrude Kraken Collagen Hydrogel 18 mg/mL.

For in vitro tissue engineering and research applications only. This product is not for use in diagnostic or therapeutic procedures.

Product Description

Kraken Collagen Hydrogel 18 mg/mL is a Type I, pH-neutral, injectable atelocollagen hydrogel designed for 3D cell culture and optimised for extrusion-based applications. Kraken Collagen Hydrogel is purified from bovine pericardium and provides a biomimetic environment that supports fundamental cellular behaviours such as adhesion, proliferation and migration, and allows for the direct encapsulation of cells without the need for a pre-neutralisation step. 

Kraken Collagen Hydrogel offers high resolution 3D tissue constructs through natural fibrillogenesis, making it suitable for a wide range of 3D biosystems applications in biomedical research, tissue engineering and Regenerative Medicine applications.

Package Contents

Each pack of Kraken Collagen Hydrogel contains one (1) x 5 mL injectable atelocollagen hydrogel 18 mg/mL and two (2) x sterile luer lock connectors.

Important Information

• Read the Instructions for Use
  
• Store at 2 - 8 °C (36 - 46°F). Do not freeze.
  
• To prevent early gelation during handling, pre-condition equipment in an ice bath.
  
• Store in cool and dry conditions away from sunlight as this may damage the collagen.
  
• This product has been sterilized. Aseptic techniques while handling is recommended.
  
• Kraken Collagen Hydrogel is best extruded at 6 - 10 °C (43 - 50 °F) through a 21 gauge needle.
  
• Ensure there are no bubbles in the solution.

Preparation of Kraken Collagen Hydrogel

Kraken Collagen Hydrogel can be used as a standalone scaffold or in combination with cells. If using a cell free suspension, follow steps 8 onward.

1. Pre-condition all syringes, needles and solutions to 2 - 8 °C (36 - 46 °F) before use. Maintain this temperature during use to ensure best results.
2. Prepare the concentrated cell suspension according to preference and place into a sterile syringe.
3. Place the sterile luer lock connector on the end of the syringe with the cell suspension. Each vial of Kraken Collagen Hydrogel comes with two (2) sterile luer lock connectors.
4. Prime the syringe by pushing the plunger until a small amount of solution is protruding out the top of the connector.
5. Uncap the collagen containing syringe and prime the syringe as in step 4.
   a) If using less than 5 mL of collagen, dispense the desired amount into a separate sterile syringe using a connector. Once dispensed, remove the connector from the original syringe and replace the screw top.
6. Join the collagen-containing syringe to the cell culture syringe. Ensure there are no air bubbles in the solution during this step, and the connector is secured to both syringes.
7. Mix the collagen and cell culture together by pushing the plungers back and forth at least 20 times until homogeneity is achieved. Vigorous mixing can lead to air bubbles and damage to the cells or collagen. Mix slowly.
8. Following mixing, ensure the entirety of the mixture is transferred to the syringe that will be used for extrusion.
   a) If using for bioprinting, transfer the mixture to a syringe or cartridge compatible with the printer.
   
9. Place on ice and allow the cell/hydrogel mixture to cool before extrusion.

Extruding Kraken Collagen Hydrogel

1. The cell-laden hydrogel can be extruded through a 21 gauge needle at 6 - 10 °C (43 - 50 °F).
2. To support the construct, we recommend using a support bath when bioprinting complex structures. Simple geometries may not require support.
3. Allow the construct to sit for approximately 5 minutes before moving to an incubator at 37 °C (98.6 °F) for 60 - 90 minutes until crosslinked.