HyStem^® Hydrogel Kits - The blank slate matrix. 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.

GS310F replaces Sigma HYS010 with a new user-friendly buffering sytem. Follow preparation instructions found in the product information sheet.

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.

Extralink is a registered trademark of Glycosan BioSystems, Inc.

Hystem is a registered trademark of BIOTIME, INC