- Hyaluronan hydrogel for stem cell culture
- 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)
HyStem is thiol-modified hyaluronic acid (HA) and is a component of the HyStem hydrogel kit. HA is a glycosaminoglycan and a major constituent of the extracellular matrix (ECM). Cells can be encapsulated in HyStem; however, cells do not attach to HyStem-only hydrogels1. For applications where cell attachment is required, HyStem can be used in conjunction with ECM proteins such as laminin, collagen, or fibronectin2. HyStem can be purchased in two formats: a 7.5 ml kit, which includes 6 – 1 ml vials of HyStem and 3 – 0.5 ml vials of Extralink, or as individual 1 ml vials.
Reconstituted HyStem remains liquid at 15 to 37°C. Without crosslinker, HyStem 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 HyStem, the gelation time is about twenty minutes with no low-temperature or low-pH steps. Diluting HyStem with phosphate-buffered saline (PBS) or cell-culture medium can increase its gelation time.
HyStem® vials come in two sizes:
- 1 mL Vial
- 5 mL Vial
For research use only
HyStem (thiol-modified sodium hyaluronate) is packaged in 1 mL and 5 mL vials. Vials are blanketed by argon and under a slight vacuum.
- Store HyStem in the original vial, unopened, at -20 °C for up to one year.
- Do not uncap the HyStem vials since they will crosslink in the presence of oxygen. Use a syringe and needle to add DG Water to the vials.
INSTRUCTIONS FOR USE
HyStem 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.
HyStem should be prepared in the following manner:
- Allow the HyStem vial to come to room temperature.
- Under aseptic conditions, using a syringe and needle, add 1 mL of DG Water to the HyStem 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.
- Typically, Extralink is used in a 1:4 volume ratio with HyStem (i.e. 1.0 mL of HyStem is crosslinked with 0.25 mL of Extralink). Note: Hydrogels made using only HyStem-HP and Extralink 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).