Product Applications Read our HyStem eBrochure Here or Click on the title of the desired protocol to learn more: HyStem Hydrogel Stiffness/Rheology Curves 2D Cell Growth on HyStem Hydrogels HyStem 3D Cell Encapsulation for Cell Delivery Applications Guide HyStem 3D Cell Encapsulation in hydrogels using 96-well plates HyStem 3D Cell Encapsulation in hydrogels using TC Inserts Enzyme Digestion of HyStem Hydrogels for Recovery of Encapsulated Cells Cell Recovery from Surface of HyStem Hydrogels HyStem Gelation Time Variation HyStem Stiffness Variation Protocol 12.5 mL Kit HyStem Stiffness Variation Protocol 7.5 mL Kit Fluorescent Labeling HyStem Hydrogels HyStem ECM Incorporation

HyStem is a 3D hydrogel kit utilizing thiolated-hyaluronic acid (HA). HA is the simplest glycosaminoglycan (a negatively charged, linear, non-sulfated polysaccharide) that provides compression strength, lubrication, and hydration within the ECM. HA regulates cell adhesion, motility, proliferation and differentiation.

HyStem is a 3D hydrogel kit utilizing thiolated-hyaluronic acid (HA). HA is the simplest glycosaminoglycan (a negatively charged, linear, non-sulfated polysaccharide) that provides compression strength, lubrication, and hydration within the ECM. HA regulates cell adhesion, motility, proliferation and differentiation.

Product Description HyStem ® Hydrogel Kits - The blank slate matrix. For first time users, we highly recommend the 2.5 mL or 7.5 mL kit size. HyStem ® hydrogels are xeno-free and completely customizable. Recommended for applications requiring specific attachment factor optimization as well as non-adherent cell culture, cellular applications. Extracellular matrix proteins can be mixed into the hydrogel and incorporated non-covalently before gelation. Alternatively, attachment peptides having an N-terminal cysteine can also be covalently linked to the matrix. The HyStem Hydrogel Kits are optimal for culturing stem cells whose natural environment is rich in hyaluronic acid (HA). HyStem hydrogels can be customized by adding extracellular matrix (ECM) proteins or cell attachment peptides into the hydrogel to provide attachment site and/or differentiation signals. They can also be varied by changing the hydrogel rigidity to match that of the native cell environment. The HyStem hydrogel system is viable for cell culture in any format, from T-flasks and tissue culture inserts to 384-well microplates. Product may be shipped ambient, but please store components at the temperature stated on the vials until ready for use. Features Dimensionality (3-D encapsulation or 2-D plating on top of hydrogel) Flexible culture format (384- through 6-well plates, tissue-culture inserts, and T-flasks) Variable amounts and type of ECM protein incorporated Gelation properties including gelation time and hydrogel stiffness Animal-free System HyStem hydrogels are xeno-free since its two components are thiol-modified hyaluronan (Glycosil ® ) and a thiol-reactive crosslinker (polyethylene glycol diacrylate, Extralink ® -Lite). The hyaluronic acid used to produce HyStem is made by a proprietary bacterial-fermentation process using bacillus subtilis (Novozymes). It is 100% free of animal-derived raw materials and no animal-derived ingredients are used in its production. Extralink (polyethylene glycol diacrylate) is made by adding acrylate groups to both ends of a polyethylene glycol (PEG) polymer. PEG is derived from petroleum and inorganic sources and contains no animal source materials. Gelation Reconstituted HyStem components remain liquid at 15 to 37°C for several hours. The hydrogel is formed when the crosslinking agent, Extralink ® -Lite (PEGDA) is added to a mixture of Glycosil ® (thiol-modified hyaluronan. Gelation occurs in about twenty minutes after all three components are mixed. No steps depend on low temperatures or low pH. Diluting the components with phosphate-buffered saline (PBS) or cell-culture medium can increase the gelation time. 3D Cell Recovery Matrix For digesting the matrix and recovering cells, we recommend using a Collagenase/Hyaluronidase mixture from Stemcell Technologies found here.