- Thiol-modified hyaluronan
- Lyophilized solid for reconstitution in DG water
- Vials are blanketed in argon and under a slight vacuum
- Tested for bacteria, endotoxins, and lactate dehydrogenase-elevating virus (LDEV)
Glycosil is thiol-modified hyaluronic acid and is a component of the Extracel® and Extracel-X® hydrogel kits. Hyaluronic acid is a major constituent of native extracellular matrix (ECM); however, some cells do not attach well to Glycosil-only hydrogels1. For these applications, Glycosil is used in conjunction with Gelin-S® or ECM proteins such as laminin, collagen, or fibronectin for most 3-D cell culture and tissue-engineering applications2. Glycosil can be purchased separately from the hydrogel kits in individual vials.
Reconstituted Glycosil remains liquid at 15 to 37°C. Without crosslinker, Glycosil will form a hydrogel via disulfide bond formation; however, the gelation time is over twenty-four hours. The rate of disulfide bond formation is highly dependent upon the solution pH and the amount of oxygen present. If Extralink® is used to crosslink the Glycosil, the gelation time is about twenty minutes with no low-temperature or low-pH steps. Diluting Glycosil with phosphate-buffered saline (PBS) or cell-culture medium can increase its gelation time.
Glycosil vials come in two sizes:
- 1 mL Vial
- 5 mL Vial
Hyaluronic Acid Source
The hyaluronic acid used to produce Glycosil® and HyStem® is made by a proprietary bacterial-fermentation process using bacillus subtilis as the host in an ISO 9001:2000 process (www.biopolymer.novozymes.com/). It is produced by Novozymes under the trade name HyaCare® and is 100% free of animal-derived raw materials and organic solvents remnants. No animal-derived ingredients are used in its production and it has very low protein levels and no exotoxins. Its production follows the European Pharmacopoeia.
For research use only
Glycosil (thiol-modified sodium hyaluronate) is packaged in 1 mL and 5 mL vials. Vials are blanketed by argon and under a slight vacuum.
- Store Glycosil in the original vial, unopened, at -20 °C for up to one year.
- Do not uncap the Glycosil vials since they will crosslink in the presence of oxygen. Use a syringe and needle to add DG Water and remove product from the vials through the septum.
INSTRUCTIONS FOR USE
Glycosil is prepared by dissolving the lyophilized solid in the DG Water (or any sterile, degassed, deionized water). When reconstituted, it will be in 1x phosphate buffered saline (PBS), pH ~7.4. The amount of DG Water used for dissolution will depend on the vial.
Glycosil should be prepared in the following manner:
- Allow the Glycosil vial to come to room temperature.
- Under aseptic conditions, using a syringe and needle, add 1 mL of DG Water to the Glycosil vial (depending on the vial size)
- Add 1.0 mL for a 1.0 mL vial
- Add 5.0 mL for a 5.0 mL vial
- Place the vial horizontally on a rocker or shaker. It will take <30 minutes for the solids to fully dissolve. Warming to 40 °C or less and/or gently vortexing will speed up dissolving time. Solutions will be clear and slightly viscous.
- Use with Extralink to form a hydrogel quickly or to allow to auto-crosslink (via disulfide bond formation) to form a hydrogel in >12 hours.
- Typically, Extralink is used in a 1:4 volume ratio with Glycosil (i.e. 1.0 mL of Glycosil is crosslinked with 0.25 mL of Extralink). Note: The gelation time will be <15 minutes (shorter than the gelation time of Glycosil + Gelin-S). Hydrogels made using only Glycosil will not support cell attachment. Use with Gelin-S for cell attachment.
Note: Product has been manufactured under aseptic conditions and tested for bacteria and fungus.
- X. Z. Shu, Y. Liu, F. Palumbo, Y. Luo, G. D. Prestwich, “In Situ Crosslinkable Hyaluronan Hydrogels for Tissue Engineering,” Biomaterials, 25, 1339-1348 (2004).
- X. Z. Shu, S. Ahmad, Y. Liu, and G. D. Prestwich, “Synthesis and Evaluation of Injectable, In Situ Crosslinkable Synthetic Extracellular Matrices (sECMs) for Tissue Engineering,” J. Biomed Mater. Res. A, 79A(4), 901-912 (2006).