Overview
The final pH, osmo, and rheological characteristics of HyStem Version 2.0 are identical to those of Version 1.0, but it is more flexible and reliable.
The fully chemically defined HyStem-HP hydrogel is perfect for cell applications where re-creating the desired microenvironment requires the slow, continuous release of growth factors. The HyStem-HP Hydrogel Kit includes thiol-modified denatured collagen (Gelin-S), thiol-modified heparin (Heprasil®), and thiol-reactive crosslinker, PEGDA (Extralink).
The extracellular matrix normally contains heparin sulfate proteoglycans, which are mimicked by the immobilized heparin in the HyStem-HP hydrogel. Proteins and heparin form an ionic bond that prevents proteolysis and speeds up the slow release of the proteins into the cell culture medium.
Compared to when growth factors are added directly to the medium, this significantly lowers the amount of growth factor required to promote cell growth or differentiation.
Image Credit: Advanced BioMatrix
Even though the product may be shipped at room temperature, it should be kept at –20 °C until needed.
Features
- Cells can be encapsulated or grown on the hydrogel surface in any format, including culture flasks, 6- to 384-well plates, or tissue culture inserts
- Hydrogels can be easily customized by the user to possess the desired stiffness and gelation time by manipulating component concentration and mixing ratios
- Growth factors can be mixed into the hydrogels prior to gelation to provide a slow growth factor release depot
- Hydrogels are suitable for animal implantation (such as angiogenesis applications and cell or drug delivery), culturing of primary cells, stem cells, and cell lines in the presence of growth factors
Applications
Gelation
- At 15 to 37 °C, the components of reconstituted HyStem-HP are still liquid
- Heprasil® (thiol-modified hyaluronan plus heparin) and Gelin-S® (thiol-modified gelatin) are combined to create the hydrogel, which is then crosslinked with Extralink® (PEGDA)
- After mixing all three ingredients, gelation occurs in about twenty minutes. Low pH or low temperatures do not really affect any steps
- The gelation time could be prolonged by diluting the ingredients with a cell culture medium or phosphate-buffered saline (PBS)
3D cell recovery matrix
- All HyStem, HyStem-C, and HyStem-HP kits can be used with the alternative crosslinker PEGSSDA for applications where cell recovery is pivotal
- With the added benefit of having internal bonds that are simple to reduce, this crosslinker offers the same benefits as Extralink
- This facilitates quick and simple recovery of single cells or clusters from the hydrogel for uses like RNA analysis or flow cytometry as opposed to time-consuming enzymatic procedures that could affect cell viability