Technical

Extracel-X™ (7.5 mL) Hydrogel Kit Product Data Sheet

For research use only

PRODUCT DESCRIPTION

The Extracel-X Hydrogel Kit is composed of Glycosil™ (thiol-modified sodium hyaluronate), Gelin-S™ (thiol-modified gelatin), Extralink™ (PEGDA, polyethylene glycol diacrylate), and degassed deionized  water (DG Water™). Solutions of Glycosil and Gelin-S form a transparent hydrogel when mixed with Extralink. All lyophilized solids are blanketed by argon and under a slight vacuum.

STORAGE

Glycosil™ and Gelin-S™

  • Store Glycosil and Gelin-S in original vials unopened at -20 °C for up to one year.
  • Do not uncap the Glycosil and Gelin-S vials since both materials disulfide crosslink in the presence of oxygen. Use a syringe and needle to add DG Water and remove product from the vials.

Extralink™

  • Store Extralink in the original vial unopened at -20 °C for up to one year.
  • Reconstituted Extralink solutions can be stored at -20 °C for ~one month.

INSTRUCTIONS FOR USE

Glycosil, Gelin-S, and Extralink solutions are prepared by dissolving the lyophilized solids in the DG Water. When reconstituted, they will be in phosophate-buffered saline (PBS), pH ~7.4. Glycosil, Gelin-S, and Extralink solutions can be used in a variety of ways depending on the desired final gel characteristics. The standard Extracel-X™ formulation with a gelation time of ~20 minutes is:

  • 1.0 mL Glycosil
  • 1.0 mL Gelin-S
  • 0.5 mL Extralink
  • Cell pellet

However, the Glycosil + Gelin-S solutions can be further diluted with medium to either increase the gelation time or to give cells more nutrients prior to formation of the hydrogel. Glycosan recommends performing a test run prior to animal experimentation to determine the optimal medium dilution for your application.

STANDARD EXTRACEL-X FORMULATION

Extracel-X hydrogels (3 x 2.5 mL = 7.5 mL) should be prepared in the following manner:

  1. Allow the Glycosil, Gelin-S, Extralink, and DG Water vials to come to room temperature.
  2. Under aseptic conditions, using a syringe and needle, add 1.0 mL of DG Water to the Glycosil vial. Repeat for the Gelin-S vial.
  3. Place both vials horizontally on a rocker or shaker. It will take <30 minutes for the solids to fully dissolve. Warming to not more than 37 °C and/or gently vortexing will speed dissolution,   Solutions will be clear and slightly viscous.
  4. Under aseptic conditions, using a syringe and needle, add 0.5 mL of DG Water to the Extralink vial. Invert several times to dissolve.
  5. As soon as possible, but within 2 hours of making the solutions, aspetically mix equal volumes of Glycosil and Gelin-S. To mix, pipette back and forth slowly to avoid trapping air bubbles.
  6. If encapsulating cells,  resuspend cell pellet in 2.0 mL of Glycosil + Gelin-S. Pipette back and forth to mix.
  7. To form the hydrogel, add Extralink to the Glycosil + Gelin-S mix in a 1:4 volume ratio (0.5 mL Extralink to 2.0 mL Glycosil + Gelin-S) and mix by pipette.
  8. Gelation will occur within ~20 minutes.

Extracel-X Gelation Time

Diluting Extracel-X with Medium

Extracel-X dilution graphExtracel-X can be further diluted with medium to either increase the gelation time or to give cells more nutrients prior to formation of the hydrogel. As Extracel-X is diluted, the hydrogel characteristics change based on the medium. Representative data is given for RPMI 1640 and DMEM (see "Extracel-X Gelation Time" graph). Note that if another medium is used, the final gelation times may be different than stated here. Contact Glycosan Technical Services (801-583-8212) if you have questions about a specific medium.

Diluted Extracel-X hydrogels (7.5 mL) can be prepared in the following manner:

  1. If frozen, allow Glycosil, Gelin-S, Extralink, and DG Water vials to thaw and come to room temperature.
  2. Under aseptic conditions, using a syringe and needle, add 1.0 mL of DG Water to the Glycosil vial. Repeat for the Gelin-S vial.
  3. Place the vial horizontally on a rocker or shaker. It will take <30 minutes for the solids to fully dissolve. Warming to not more than 40 °C and/or gently vortexing will speed dissolution. The solution will be clear and slightly viscous.
  4. Under aseptic conditions, using a syringe and needle, add 0.5 mL of DG Water to the Extralink vial. Invert several times to dissolve.
  5. As soon as possible, but within 2 hours of making the solutions, aseptically mix equal volumes of Glycosil and Gelin-S. To mix, pipette back and forth slowly to avoid trapping air bubbles.
  6. Based on the desired dilution, add the following volumes of medium + cells to the 10.0 mL of Glycosil/Gelin-S mix:
    Dilution, % 10 20 30 40 50
    Medium + cells, mL 0.22 0.50 0.86 1.33 2.00
  7. Pipette back and forth to mix.
  8. To form the hydrogel, add Extralink to the Glycosil + Gelin-S mix in a 1:4 volume ratio (0.5 mL Extralink to 2.0 mL Glycosil + Gelin-S + media + cells).
  9. See "Extracel-X Gelation Time" graph for the expected gelation time.

Note: Each kit component has been manufactured under aseptic conditions and tested for bacteria and fungus.

For more information about the Extracel-X Hydrogel Kit go to www.glycosan.com.